High penetration drugs and their compositions thereof for treatment of parkinson diseases

ABSTRACT

One aspect of the invention provides a composition of novel high penetration compositions (HPC) or a high penetration prodrug (HPP) for treatment of Parkinson&#39;s disease. The HPCs/HPPs are capable of being converted to parent active drugs or drug metabolites after crossing the biological barrier and thus can render treatments for the conditions that the parent drugs or metabolites can. Additionally, the HPPs are capable of reaching areas that parent drugs may not be able to access or to render a sufficient concentration at the target areas and therefore render novel treatments. The HPCs/HPPs can be administered to a subject through various administration routes, e.g., locally delivered to an action site of a condition with a high concentration or systematically administered to a biological subject and enter the general circulation with a faster rate.

PRIORITY CLAIM

This application is a continuation of International Patent ApplicationNo. PCT/CN/13/72728, filed Mar. 15, 2013, which is incorporated byreference herein in its entirety, including the drawings.

FIELD OF THE INVENTION

The invention relates to the field of pharmaceutical compositions. Morespecifically, one aspect of the invention relates to pharmaceuticalcompositions capable of penetrating one or more biological barriers andmethods of using the pharmaceutical compositions for preventing and/ortreating Parkinson's disease and/or Parkinsonian syndromes in a subject.

BACKGROUND OF THE INVENTION

Parkinson's disease (PD) is a neurological disorder characterized by adegeneration of dopamine neurons in the substantia nigra and a loss ofdopamine in the putamen. It is described as a motor disease, but it alsoproduces cognitive and behavioral symptoms. Parkinson's disease mayresult from the death of dopamine-generation cells in the substatianigra, a region of the midbrain. The cause of this cell death may beunknown (primary parkisonism) or known (secondary parkinsonism).Parkinson's disease may cause many symptoms in a wide variety offunctions including movement, attention and learning. The primarysymptoms of Parkinson's disease result from reduced activity ofdopamine-secreting cells caused by cell death in the pars compactaregion of the substantia nigra. A particular conceptual model of themotor circuit and its alteration with PD has been of great influencesince 1980, although some limitations have been pointed out which haveled to modifications [Obeso J A, Rodrìguez-Oroz M C, Benitez-Temino B,et al. (2008). “Functional organization of the basal ganglia:therapeutic implications for Parkinson's disease”. Mov. Disord. 23(Suppl 3): S548-59]. In this model, the basal ganglia normally exert aconstant inhibitory influence on a wide range of motor systems,preventing them from becoming active at inappropriate times. When adecision is made to perform a particular action, inhibition is reducedfor the required motor system, thereby releasing it for activation.Dopamine acts to facilitate this release of inhibition, so high levelsof dopamine function tend to promote motor activity, while low levels ofdopamine function, such as occur in PD, demand greater exertions ofeffort for any given movement. Thus the net effect of dopamine depletionis to produce hypokinesia [Obeso J A, Rodriguez-Oroz M C, Benitez-TeminoB, et al. (2008). “Functional organization of the basal ganglia:therapeutic implications for Parkinson's disease”. Mov. Disord. 23(Suppl 3): S548-59].

The main families of drugs known for treating motor symptoms arelevodopa (usually combined with a dopa decarboxylase inhibitor or COMTinhibitor), dopamine agonists and MAO-B inhibitors [The NationalCollaborating Centre for Chronic Conditions, ed. (2006). “Symptomaticpharmacological therapy in Parkinson's disease”, Parkinson's Disease.London: Royal College of Physicians. pp. 59-100]. Levodopa is well knownto improve motor symptoms but its effects in cognitive and behavioralsymptoms are more complex [Cools R (2006). “Dopaminergic modulation ofcognitive function-implications for L-DOPA treatment in Parkinson'sdisease”. Neurosci Biobehav Rev 30 (1): 1-23]. Levodopa preparationscontribute to the development of motor complications in PD. Thesecomprise abnormal involuntary movements or dyskinesias, such asathetosis and dystonia, along with response fluctuations in which peopleexperience “wearing off” of the drug's effects and/or unpredictableswitching between the “on” and the “off” state. Levodopa may also causenausea, vomiting, gastrointestinal bleeding, dyskinesia at peak dose,and end-of-dose deterioration of function.

Thus, there is a need to develop better treatments to address thoseproblems.

BACKGROUND OF THE INVENTION

One aspect of the invention relates to HPPs of levodopa and/or dopamine.

Another aspect of the invention relates to pharmaceutical compositionscomprising one or more HPPs disclosed herein. In certain embodiments,the pharmaceutical composition comprises one or more high penetrationprodrugs of NSAID and one or more high penetration prodrugs of dopamineand/or levodopa.

where the parent drug(s) of the HPPs may be the same or different, andmay be levodopa, dopamine, aspirin, ibuprofen, and/or other NSAIDs asdisclosed herein.

Another aspect of the invention relates to a method of using acomposition of the invention in penetrating one or more biologicalbarriers in a biological subject.

Another aspect of the invention relates to a method of using acomposition of the invention, or a pharmaceutical composition thereof intreating a condition in a biological subject.

Another aspect of the invention relates to a method of using one or moreHPPs or a pharmaceutical composition thereof in treating Parkinson'sdisease and/or related conditions in a biological subject or subject byadministrating the one or more HPPs or a pharmaceutical compositionthereof to the biological subject or subject.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1. The staying time results in Totarod test after 1-3 weektreatment (n=12) (Example 31).

FIG. 2. The falling speed results in Totarod test after 1-3 weektreatment (n=12) (Example 31).

FIG. 3. The staying time results in Totarod test after 4 weeks treatment(n=12) (Example 32).

FIG. 4. The falling speed results in Totarod test after 4 weekstreatment (n=12) (Example 32).

FIG. 5. The staying time results in Totarod test after 4 weeks treatment(n=12) (Example 33).

FIG. 6. The falling speed results in Totarod test after 4 weekstreatment (n=12) (Example 33).

FIG. 7. The staying time results in Totarod test after 4 weeks treatment(n=12) (Example 34).

FIG. 8. The falling speed results in Totarod test after 4 weekstreatment (n=12) (Example 34).

FIG. 9. The falling speed results in Totarod test after 4 weekstreatment (n=12) (Example 35).

FIG. 10. The falling speed results in Totarod test after 4 weekstreatment (n=12) (Example 35).

FIG. 11. The staying time results in Totarod test after 4 weekstreatment (n=12) (Example 36).

FIG. 12. The falling speed results in Totarod test after 4 weekstreatment (n=12) (Example 36).

FIG. 13. The staying time results in Totarod test after 4 weekstreatment (n=12) (Example 37).

FIG. 14. The falling speed results in Totarod test after 4 weekstreatment (n=12) (Example 37).

FIG. 15. The staying time results in Totarod test after 4 weekstreatment (n=12) (Example 38).

FIG. 16. The falling speed results in Totarod test after 4 weekstreatment (n=12) (Example 38).

DETAILED DESCRIPTION OF THE INVENTION I. High Penetration Prodrugs ofLevodopa

One aspect of the invention relates to a high penetration prodrug oflevodopa having a structure selected from the group consisting ofStructure Pro-L-Dopa-1, Structure Pro-L-Dopa-2, Structure Pro-L-Dopa-3,Structure Pro-L-Dopa-4 and Structure Pro-L-Dopa-5:

including stereoisomers and pharmaceutically acceptable salts thereof,wherein:

W is selected from the group consisting of H, substituted andunsubstituted alkyl, substituted and unsubstituted cycloalkyl,substituted and unsubstituted heterocycloalkyl, substituted andunsubstituted alkyloxy, substituted and unsubstituted alkenyl,substituted and unsubstituted alkynyl, substituted and unsubstitutedaryl, substituted and unsubstituted heteroaryl, the protonatable aminegroup, pharmaceutically acceptable substituted and unsubstituted aminegroups, Structure W-1, Structure W-2, Structure W-3, Structure W-4,Structure W-5, Structure W-6, Structure W-7, Structure W-8, StructureW-9, Structure W-10, Structure W-11, and Structure W-12:

HA is selected from the group consisting of nothing, andpharmaceutically acceptable acid, e.g. hydrochloride hydrobromide,hydroiodide, nitric acid, sulfic acid, bisulfic acid, phosphoric acid,phosphorous acid, phosphonic acid, isonicotinic acid, acetic acid,lactic acid, salicylic acid, citric acid, tartaric acid, pantothenicacid, bitartaric acid, ascorbic acid, succinic acid, maleic acid,gentisinic acid, fumaric acid, gluconic acid, glucaronic acid, saccharicacid, formic acid, benzoic acid, glutamic acid, methanesulfonic acid,ethanesulfonic acid, benzensulfonic acid, p-toluenesulfonic acid andpamoic acid;

R₁ and R₂ are independently selected from the group consisting of H,substituted and unsubstituted alkyl, substituted and unsubstitutedcycloalkyl, substituted and unsubstituted heterocycloalkyl, substitutedand unsubstituted alkyloxyl, substituted and unsubstituted alkenyl,substituted and unsubstituted alkynyl, substituted and unsubstitutedaryl and substituted and unsubstituted heteroaryl residues;

R₁₁-R₁₅ are independently selected from the group consisting of nothing,H, CH₂C(═O)OR₁₁, substituted and unsubstituted alkyl, substituted andunsubstituted cycloalkyl, substituted and unsubstitutedheterocycloalkyl, substituted and unsubstituted alkoxyl, substituted andunsubstituted perfluoroalkyl, substituted and unsubstituted alkylhalide, substituted and unsubstituted alkenyl, substituted andunsubstituted alkynyl, substituted and unsubstituted aryl, andsubstituted and unsubstituted heteroaryl;

X is selected from the group consisting of O, S, and NR₃;

X₂ is selected from the group consisting of nothing, O, S, NR₃, CHR₃—O,CHR₃—S, CHR₃—O, O—CHR₃—O, O—CHR₃—S, S—CHR₃—O, and S—CHR₃—S;

X₃ is selected from the group consisting of nothing, C═O, C═S, C(═O)—O,O, S, NR₃, C(═O)—O—CHR₃—O, C(═O)—O—CHR₃—S, C(═O)—S—CHR₃—O, andC(═O)—S—CHR₃—S:

Y₁ is selected from the group consisting of R₃C(═O), R₃O—C(═O), andR₃S—C(═O);

Y₂ is selected from the group consisting of R₃C(═O), R₃O—C(═O), andR₃S—C(═O);

Y₃ is selected from the group consisting of R₃, OR₃, SR₃, NR₃R₄,O—CHR₃—OR₄, O—CHR₃—SR₄, and S—CHR₃—OR₄;

n and m is selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7,and 8.

R_(c) is selected from the group consisting of nothing, CH₂C(═O)OR₆,substituted and unsubstituted alkyl, substituted and unsubstitutedcycloalkyl, substituted and unsubstituted heterocycloalkyl, substitutedand unsubstituted alkoxyl, substituted and unsubstituted perfluoroalkyl,substituted and unsubstituted alkyl halide, substituted andunsubstituted alkenyl, substituted and unsubstituted alkynyl,substituted and unsubstituted aryl, and substituted and unsubstitutedheteroaryl, wherein any CH₂ in R may be further replaced with O, S, P,or NR₆;

R is selected from the group consisting of nothing, CH₂C(═O)OR₆,substituted and unsubstituted alkyl, substituted and unsubstitutedcycloalkyl, substituted and unsubstituted heterocycloalkyl, substitutedand unsubstituted alkoxyl, substituted and unsubstituted perfluoroalkyl,substituted and unsubstituted alkyl halide, substituted andunsubstituted alkenyl, substituted and unsubstituted alkynyl,substituted and unsubstituted aryl, and substituted and unsubstitutedheteroaryl, wherein any CH₂ in R may be further replaced with O, S, P,or NR₆;

R₃ and R₄ are independently selected from the group consisting ofnothing, CH₂C(═O)OR₆, substituted and unsubstituted alkyl, substitutedand unsubstituted cycloalkyl, substituted and unsubstitutedheterocycloalkyl, substituted and unsubstituted alkoxyl, substituted andunsubstituted perfluoroalkyl, substituted and unsubstituted alkylhalide, substituted and unsubstituted alkenyl, substituted andunsubstituted alkynyl, substituted and unsubstituted aryl, andsubstituted and unsubstituted heteroaryl, wherein any CH₂ in R may befurther replaced with O, S, P, or NR₆;

Each R₆ is independently selected from the group consisting of H, F, Cl,Br, I, Na⁺, K⁺, C(═O)R₅, 2-oxo-1-imidazolidinyl, phenyl, 5-indanyl,2,3-dihydro-1H-inden-5-yl, 4-hydroxy-1,5-naphthyridin-3-yl, substitutedand unsubstituted alkyl, substituted and unsubstituted cycloalkyl,substituted and unsubstituted heterocycloalkyl, substituted andunsubstituted alkenyl, substituted and unsubstituted alkynyl,substituted and unsubstituted alkyloxyl, substituted and unsubstitutedcycloalkyloxyl, substituted and unsubstituted aryl, substituted andunsubstituted heteroaryl, —C(═O)—W, -L₁-L₄-L₂-W, and W;

each R₅ is independently selected from the group consisting of H,C(═O)NH₂, CH₂CH₂OR₆, CH₂CH₂N(CH₃)₂, CH₂CH₂N(CH₂CH₃)₂, Cl, F, Br, I,substituted and unsubstituted alkyl, substituted and unsubstitutedcycloalkyl, substituted and unsubstituted heterocycloalkyl, substitutedand unsubstituted alkyloxyl, substituted and unsubstitutedcycloalkyloxyl, substituted and unsubstituted aryl, substituted andunsubstituted heteroaryl, substituted and unsubstituted alkylcarbonyl,substituted and unsubstituted alkylamino, —C(═O)—W, L₁-L₄-L₂-W, and W;

L₁ is selected from the group consisting of nothing, O, S, —O-L₃-,—S-L₃-, —N(L₃)-, —N(L₃)-CH₂—O, —N(L₃)-CH₂—N(L₅)-, —O—CH₂—O—,—O—CH(L₃)-O, and —S—CH(L₃)-O—;

L₂ is selected from the group consisting of nothing, O, S, —O-L₃-,—S-L₃-, —N(L₃)-, —N(L₃)-CH₂—O, —N(L₃)-CH₂—N(L₅)-, —O—CH₂—O—,—O—CH(L₃)-O, —S—CH(L₃)-O—, —O-L₃-, —N-L₃-, —S-L₃-, —N(L₃)-L₅- and L₃;

L₄ is selected from the group consisting of nothing, C═O, C═S,

for each L₁, L₂, and L₄, each L₃ and L₅ is independently selected fromthe group consisting of nothing, H, CH₂C(═O)OL₆, substituted andunsubstituted alkyl, substituted and unsubstituted cycloalkyl,substituted and unsubstituted heterocycloalkyl, substituted andunsubstituted aryl, substituted and unsubstituted heteroaryl,substituted and unsubstituted alkoxyl, substituted and unsubstitutedalkylthio, substituted and unsubstituted alkylamino, substituted andunsubstituted perfluoroalkyl, and substituted and unsubstituted alkylhalide, wherein any carbon or hydrogen may be further independentlyreplaced with O, S, P, or NL₃;

each L₆ is independently selected from the group consisting of H, OH,Cl, F, Br, I, substituted and unsubstituted alkyl, substituted andunsubstituted cycloalkyl, and substituted and unsubstitutedheterocycloalkyl, substituted and unsubstituted aryl, substituted andunsubstituted heteroaryl, substituted and unsubstituted alkoxyl,substituted and unsubstituted alkylthio, substituted and unsubstitutedalkylamino, substituted and unsubstituted perfluoroalkyl, andsubstituted and unsubstituted alkyl halide, wherein any carbon orhydrogen may be further independently replaced with O, S, N, P(O)OL₆,CH═CH, C≡C, CHL₆, CL₆L₇, aryl, heteroaryl, or cyclic groups;

each L₇ is independently selected from the group consisting of H, OH,Cl, F, Br, I, substituted and unsubstituted alkyl, substituted andunsubstituted cycloalkyl, and substituted and unsubstitutedheterocycloalkyl, substituted and unsubstituted aryl, substituted andunsubstituted heteroaryl, substituted and unsubstituted alkoxyl,substituted and unsubstituted alkylthio, substituted and unsubstitutedalkylamino, substituted and unsubstituted perfluoroalkyl, andsubstituted and unsubstituted alkyl halide, wherein any carbon orhydrogen may be further independently replaced with O, S, N, P(O)OL₆,CH═CH, C≡C, CHL₆, CL₆L₇, aryl, heteroaryl, or cyclic groups; and

any CH₂ groups may be replaced with O, S, or NH.

II. High Penetration Prodrugs of Dopamine

Another aspect of the invention relates to a high penetration pro-drugof dopamine comprises a structure selected from the group consisting ofStructure Pro-dopamine-1, Structure Pro-dopamine-2, StructurePro-dopamine-3, and Structure Pro-dopamine-4, and StructurePro-dopamine-5:

including stereoisomers and pharmaceutically acceptable salts thereof,wherein W, HA, X, X₂, X₃, Y₁, Y₂, Y₃, n, m, R_(c), R, R₁, R₂, R₃, R₄,R₅, R₆, R₁₁-R₁₅, L₁, L₂, and L₄ are defined the same as supra.

III. Pharmaceutical Compositions Comprising a High Penetration Prodrugof NSAID and One or More High Penetration Prodrug of Dopamine and/orLevodopa

Another aspect of the invention relates to a pharmaceutical compositioncomprising one or more high penetration prodrugs of NSAID and one ormore high penetration prodrugs of dopamine and/or levodopa.

A high penetration pro-drug of NSAID comprises a structure selected fromthe group consisting of Structure NSAID-1, Structure NSAID-2, StructureNSAID-3, Structure NSAID-4, Structure NSAID-5, Structure NSAID-6,Structure NSAID-7, Structure NSAID-8, Structure NSAID-9, StructureNSAID-10, Structure NSAID-11, Structure NSAID-12, and StructureNSAID-13:

including stereoisomers and pharmaceutically acceptable salts thereof,wherein:

Aryl- is selected from the group consisting of Aryl-1, Aryl-2, Aryl-3,Aryl-4, Aryl-5, Aryl-6, Aryl-7, Aryl-8, Aryl-9, Aryl-10, Aryl-11,Aryl-12, Aryl-13, Aryl-14, Aryl-15, Aryl-16, Aryl-17, Aryl-18, Aryl-19,Aryl-20, Aryl-21, Aryl-22, Aryl-23, Aryl-24, Aryl-25, Aryl-26, Aryl-27,Aryl-28, Aryl-29, Aryl-30, Aryl-31, Aryl-32, Aryl-33, Aryl-34, Aryl-35,Aryl-36, Aryl-37, Aryl-38, Aryl-39, Aryl-40, Aryl-41, Aryl-42, Aryl-43,Aryl-44, Aryl-45, Aryl-46, Aryl-47, Aryl-48, Aryl-49, Aryl-50, Aryl-51,Aryl-52, Aryl-53, Aryl-54, Aryl-55, Aryl-56, Aryl-57, Aryl-58, Aryl-59,Aryl-60, Aryl-61, Aryl-62, Aryl-63, Aryl-64, Aryl-65, Aryl-66, Aryl-67,Aryl-68, Aryl-69, Aryl-70, and Aryl-71:

W, HA, X, X₂, X₃, Y₁, Y₂, Y₃, n, m, R_(c), R, R₁, R₂, R₃, R₄, R₅, R₆,R₁₁-R₁₅, L₁, L₂, and L₄, are defined the same as supra;

X₆ and X₈ are independently selected from the group consisting ofnothing, C(═O), C(═S), OC(═O), OC(═S), CH₂, CH, S, O and NR₅;

Y₄, Y₅, Y₆, Y₇, Y₈, and Y₉ are independently selected from the groupconsisting of H, OH, OW, OC(═O)W, L₁-L₄-L₂-W, OC(═O)CH₃, CH₃, C₂H₅,C₃H₇, C₄H₉, R₆, SO₃R₆, CH₂OR₆, CH₂OC(═O)R₆, CH₂C(═O)OR₈, OCH₃, OC₂H₅,OR₆, CH₃SO₂, R₆SO₂, CH₃SO₃, R₆SO₃, NO₂, ON, CF₃, OCF₃,CH₂(CH₂)_(n)NR₅R₆, CH₂(CH₂)_(n)OR₆, CH(C(═O)NH₂)NHR₆, CH₂C(═O)NH₂, F,Br, I, Cl, CH═CHC(═O)NHCH₂C(═O)OW, CH═CHC(═O)NHCH₂L₁-L₄-L₂-W,NR₈C(═O)R₅, SO₂NR₅R₈, C(═O)R₅, SR₅, R₆OOCCH(NHR₇)(CH₂)_(n)C(═O)NH—,R₆OOCCH(NHR₇)(CH₂)_(n)SC(═O)NH—, CF₃SCH₂C(═O)NH—, CF₃CH₂C(═O)NH—,CHF₂SCH₂C(═O)NH—, CH₂FSCH₂C(═O)NH—, NH₂C(═O)CHFS—CH₂C(═O)NH—,R₇NHCH(C(═O)OW)CH₂SCH₂C(═O)NH—, R₇NHCH(L₁-L₄-L₂-W)CH₂SCH₂C(═O)NH—,CNCH₂SCH₂C(═O)NH—, CH₃(CH₂)_(n)C(═O)NH—, R₇N═CHNR₇CH₂CH₂S—,R₇N═C(NHR₇)NHC(═O)—, R₇N═C(NHR₇)NHC(═O)CH₂, CH₃C(Cl)═CHCH₂SCH₂C(═O)NH—,(CH₃)₂C(OR₆)—, CNCH₂C(═O)NH—, CNCH₂CH₂S—, R₇HN═CH(NR₇)CH₂CH₂S—,CH₂═CHCH₂SCH₂C(═O)NH—, CH₃CH(OH)—, CH₃CH(OR₈)—, CH₃CH(Y₁)—, (CH₃)₂CH—,CH₃CH₂—, CH₃(CH₂)_(n)CH═CH(CH₂)_(m)C(═O)NH—, substituted andunsubstituted perfluoroalkyl, substituted and unsubstituted alkoxyl,substituted and unsubstituted alkylthio, substituted and unsubstitutedalkylamino, substituted and unsubstituted perfluoroalkyl, substitutedand unsubstituted alkyl halide and substituted and unsubstitutedalkylcarbonyl;

each R₇ is independently selected from the group consisting of H, F, Cl,Br, I, CH₃NHC(═O)CH₂CH(NHR₈)C(═O), R₅N═C(NHR₆)NHC(═O)—, C(═O)CH₃,C(═O)R₆, PO(OR₅)OR₆, substituted and unsubstituted alkyl, substitutedand unsubstituted cycloalkyl, substituted and unsubstitutedheterocycloalkyl, substituted and unsubstituted alkyloxyl, substitutedand unsubstituted alkenyl, substituted and unsubstituted alkynyl,substituted and unsubstituted aryl, substituted and unsubstitutedheteroaryl, substituted and unsubstituted alkylcarbonyl, substituted andunsubstituted alkylamino, L₁-L₄-L₂-W, and C—(═O)—W; and

each R₈ is independently selected from the group consisting of H, F, Cl,Br, I, CH₃, C₂H₅, CF₃, CH₂CH₂F, CH₂CH₂Cl, CH₂CH₂Br, CH₂CH₂I, CH₂CHF₂,CH₂CF₃, CH₂F, CH₂Cl, CH₂Br, CH₂I, CH₂NR₆R₇, CH(NHR₇)CH₂C(═O)NH₂, C₃H₇,C₄H₉, C₅H₁₁, R₆, C(═O)R₆, C(═O)NH₂, CH₂C(═O)NH₂, CH₂OC(═O)NH₂,PO(OR₅)OR₆, C(CH₃)₂C(═O)OR₆, CH(CH₃)C(═O)OR₆, CH₂C(═O)OR₆, C(═O)—W,L₁-L₄-L₂-W, W, substituted and unsubstituted perfluoroalkyl, substitutedand unsubstituted alkyl, substituted and unsubstituted cycloalkyl,substituted and unsubstituted heterocycloalkyl, substituted andunsubstituted alkoxyl, substituted and unsubstituted alkylamino,substituted and unsubstituted perfluoroalkyl, substituted andunsubstituted alkyl halide and substituted and unsubstitutedalkylcarbonyl.

As used herein, the term “pharmaceutically acceptable salt” means thosesalts of compounds of the invention that are safe for application in asubject. Pharmaceutically acceptable salts include salts of acidic orbasic groups present in compounds of the invention. Pharmaceuticallyacceptable acid addition salts include, but are not limited to,hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate,phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate,citrate, tartrate, pantothenate, bitartrate, ascorbate, succinate,maleate, gentisinate, fumarate, gluconate, glucaronate, saccharate,formate, benzoate, glutamate, methanesulfonate, ethanesulfonate,benzensulfonate, p-toluenesulfonate and pamoate (i.e.,1,11-methylene-bis-(2-hydroxy-3-naphthoate)) salts. Certain compounds ofthe invention can form pharmaceutically acceptable salts with variousamino acids. Suitable base salts include, but are not limited to,aluminum, calcium, lithium, magnesium, potassium, sodium, zinc, anddiethanolamine salts. For a review on pharmaceutically acceptable saltssee BERGE ET AL., 66 J. PHARM. SCI. 1-19 (1977), incorporated herein byreference.

As used herein, unless specified otherwise, the term “alkyl” means abranched or unbranched, saturated or unsaturated, monovalent ormultivalent hydrocarbon group, including saturated alkyl groups, alkenylgroups and alkynyl groups. Examples of alkyl include, but are notlimited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl,pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, ethenyl,propenyl, butenyl, isobutenyl, pentenyl, hexenyl, heptenyl, octenyl,nonenyl, decenyl, undecenyl, dodecenyl, ethynyl, propynyl, butynyl,isobutynyl, pentynyl, hexynyl, heptynyl, octynyl, nonynyl, decynyl,undecynyl, dodecynyl, methylene, ethylene, propylene, isopropylene,butylene, isobutylene, t-butylene, pentylene, hexylene, heptylene,octylene, nonylene, decylene, undecylene and dodecylene. In certainembodiments, the hydrocarbon group contains 1 to 30 carbons. In certainembodiments, the hydrocarbon group contains 1 to 20 carbons. In certainembodiments, the hydrocarbon group contains 1 to 12 carbons.

As used herein, unless specified otherwise, the term “cycloalkyl” meansan alkyl which contains at least one ring and no aromatic rings.Examples of cycloalkyl include, but are not limited to, cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl,cyclononyl, cyclodecyl, cycloundecyl and cyclododecyl. In certainembodiments, the hydrocarbon chain contains 1 to 30 carbons. In certainembodiments, the hydrocarbon group contains 1 to 20 carbons. In certainembodiments, the hydrocarbon group contains 1 to 12 carbons.

As used herein, unless specified otherwise, the term “heterocycloalkyl”means a cycloalkyl wherein at least one ring atom is a non-carbon atom.Examples of the non-carbon ring atom include, but are not limited to, S,O and N.

As used herein, unless specified otherwise, the term “alkoxyl” means analkyl, cycloalkyl or heterocycloalkyl, which contains one or more oxygenatoms. Examples of alkoxyl include, but are not limited to, —CH₂—OH,—OCH₃, —O—R_(e), —R_(e)—OH, —R_(e1)—O—R_(e2)—, wherein R_(e), R_(e1) andR_(e2) can be the same or different alkyl, cycloalkyl orheterocycloalkyl.

As used herein, unless specified otherwise, the term “alkyl halide”means an alkyl, cycloalkyl or heterocycloalkyl, which contains one ormore halogen atoms, wherein the halogen atoms can be the same ordifferent. The term “halogen” means fluorine, chlorine, bromine oriodine. Examples of alkyl halide include, but are not limited to,—R_(e)—F, —R_(e)—Cl, —R_(e)—Br, —R_(e)—I, —R_(e)(F)—, —R_(e)(Cl)—,—R_(e)(Br)— and —R_(e)(I)—, wherein R_(e) is an alkyl, cycloalkyl orheterocycloalkyl.

As used herein, unless specified otherwise, the term “alkylthio” meansan alkyl, cycloalkyl or heterocycloalkyl, which contains one or moresulfur atoms. Examples of alkylthio include, but are not limited to,—CH₂—SH, —SCH₃, —S—R_(e), —R_(e)—SH, —R_(e1)—S—R_(e2)—, wherein R_(e),R_(e1) and R_(e2) are the same or different alkyl, cycloalkyl orheterocycloalkyl.

As used herein, unless specified otherwise, the term “alkylamino” meansan alkyl, cycloalkyl or heterocycloalkyl, which contains one or morenitrogen atoms. Examples of alkylamino include, but are not limited to,—CH₂—NH, —NCH₃, —N(R_(e1))—R_(e2), —N—R_(e), —R_(e)—NH₂,—R_(e1)—N—R_(e2) and —R_(e)—N(R_(e1))—R_(e2) wherein R_(e), R_(e1) andR_(e2) are the same or different alkyl, cycloalkyl or heterocycloalkyl.

As used herein, unless specified otherwise, the term “alkylcarbonyl”means an alkyl, cycloalkyl or heterocycloalkyl, which contains one ormore carbonyl groups. Examples of alkylcarbonyl group include, but arenot limited to, aldehyde group (—R_(e)—C(O)—H), ketone group(—R_(e)—C(O)—R_(e1)), carboxylic acid group (R_(e)—C(═O)OH), ester group(—R_(e)—C(═O)O—R_(e1)), carboxamide, (—R_(e)—C(═O)O—N(R_(e1))R_(e2)),enone group (—R_(e)—C(O)—C(R_(e1))═C(R_(e2))R_(e3)), acyl halide group(—R_(e)—C(O)—X_(h)) and acid anhydride group(—R_(e)—C(O)—O—C(O)—R_(e1)), wherein R_(e), R_(e1), R_(e2) and R_(e3)are the same or different alkyl, cycloalkyl, or heterocycloalkyl; andX_(h) is a halogen.

As used herein, unless specified otherwise, the term “perfluoroalkyl”means an alkyl, cycloalkyl or heterocycloalkyl, which contains one ormore fluoro group, including, without limitation, perfluoromethyl,perfluoroethyl, perfluoropropyl.

As used herein, unless specified otherwise, the term “aryl” means achemical structure comprising one or more aromatic rings. In certainembodiments, the ring atoms are all carbon. In certain embodiments, oneor more ring atoms are non-carbon, e.g. oxygen, nitrogen, or sulfur(“heteroaryl”). Examples of aryl include, without limitation, phenyl,benzyl, naphthalenyl, anthracenyl, pyridyl, quinoyl, isoquinoyl,pyrazinyl, quinoxalinyl, acridinyl, pyrimidinyl, quinazolinyl,pyridazinyl, cinnolinyl, imidazolyl, benzimidazolyl, purinyl, indolyl,furanyl, benzofuranyl, isobenzofuranyl, pyrrolyl, indolyl, isoindolyl,thiophenyl, benzothiophenyl, pyrazolyl, indazolyl, oxazolyl,benzoxazolyl, isoxazolyl, benzisoxazolyl, thiaxolyl, quanidino andbenzothiazolyl.

In certain embodiments, the pharmaceutical composition comprises atleast one HPP of a parent drug that can be used to treat Parkinson'sdisease or a related compound thereof, and a pharmaceutically acceptablecarrier.

In certain embodiments, a pharmaceutical composition may comprise morethan one HPPs of the same or different parent drugs. The differentparent drugs can belong to the same or different categories of drugsthat are used to treat Parkinson's disease. For example, apharmaceutical composition may comprise HPPs of parent drugs or relatedcompounds thereof, wherein the parent drugs are selected from the groupconsisting of Levodopa, dopamine, aspirin and other nonsteroidalanti-inflammatory drugs (NSAIDs), and any combinations thereof.

A pharmaceutical composition may further comprise water.

A pharmaceutical composition may further comprise an alcohols (e.g.,ethanol, glycerol, isopropanol, octanol, etc.).

In certain embodiments, a pharmaceutical composition comprises HPPs ofparent drugs or related compounds thereof, wherein at least one of theparent drugs is levodopa (e.g. HPPs comprising one or more structures ofstructure Pro-L-dopa-1, Pro-L-dopa-2, Pro-L-dopa-3, Pro-L-dopa-4, and/orPro-L-dopa-5); dopamine (e.g. HPPs comprising one or more structures ofstructure Pro-dopamine-1, Pro-dopamine-2, Pro-dopamine-3,Pro-dopamine-4, and/or Structure Pro-dopamine-5); and at least one ofthe parent drug is aspirin or other anti-inflammatory drugs (e.g. HPPscomprising one or more structures selected from the group consisting ofStructure NSAID-1, Structure NSAID-2, Structure NSAID-3, StructureNSAID-4, Structure NSAID-5, Structure NSAID-6, Structure NSAID-7,NSAID-8, Structure NSAID-9, Structure NSAID-10, Structure NSAID-11,Structure NSAID-12, and Structure NSAID-13.)

In certain embodiments, a pharmaceutical composition comprises4-(2-(2-(methylamino)acetamido)ethyl)-1,2,-phenylene dibenzoatehydrochloride, 4-(2-(2-(methylamino)acetamido)ethyl)-1,2,-phenylenedibenzoate acetic acid,4-(6-methyl-4,8-dioxo-5,7-dioxa-2,9-diazadecan-11-yl)-1,2,-phenylenedibenzoate hydrobromide,4-(2-(2-amino-3-phenylpropanamido)ethyl)-1,2,-phenylene dibenzoatehydrochloride,4-(2-(((1-((pyrrolidine-2-carbonyl)oxy)ethoxy)amino)ethyl)-1,2,-phenylenedibenzoate hydrochloride,4-(2-piperidine-4-carboxamido)ethyl)-1,2,-phenylenebis(2-ethylbutanoate) hydrochloride,4-(2-((((-octahydro-1H-quinolizin-3-yl)oxy)carbonyl)amino)ethyl)-1,2,-phenylenebis(2-ethylbutanoate)acetate,1-(((2-(4-amino-2,5-dioxo-2,3,4,5-tetrahydrobenzo[b][1,4]dioxocin-8-yl)ethyl)carbamoyl)oxy)ethylisobutyrate hydrofluoride,1-(((2-(3-amino-2,5-dioxo-2,3,4,5-tetrahydrobenzo[b][1,4]dioxocin-8-yl)ethyl)carbamoyl)oxy)ethylisobutyrate hydrofluoride,(-(2-((ethoxycarbonyl)amino)ethyl)-2-(2-methylamino)acetoxy)phenylbenzoate hydrochloride,5-(2-((ethoxycarbonyl)amino)ethyl)-2-(2-methylamino)acetoxy)phenylbenzoate hydrochloride, 4-(2-aminoethyl)-1,2-phenylene dibenzoatehydrochloride, (S)-4-(2-amino-3-isopropoxy-3-oxopropyl)-1,2-phenylenedibenzoate hydrochloride,(S)-4-(2-amino-3-(heptan-4-yloxy)-3-oxopropyl)-1,2-phenylene dibenzoatehydrochloride, (S)-4-(2-amino-3-isopropoxy-3-oxopropyl)-1,2-phenylenedipentanoate hydrochloride,(S)-4-(2-amino-3-ethoxy-3-oxopropyl)-1,2-phenylene diacetatehydrochloride,(S)-4-(2-amino-3-oxo-3-(pentan-3-yloxy)propyl)-1,2-phenylenebis(2-methylpropanoate) hydrobromide,(S)-4-(2-aminoacetamido)-3-isopropoxy-3-oxopropyl)-1,2-phenylenedibenzoate hydrochloride,4-((2S)-3-oxo-3-(pentan-3-yloxy)-2-(pyrrolidine-2-carboxamido)propyl)-1,2-phenylenedibenzoate hydrofluoride,(S)-4-(3-isopropoxy-3-oxo-2-(piperidine-4-carboxamido)propyl)-1,2-phenylenedibenzoate hydrochloride,4-((2S)-3-isopropoxy-3-2-(octahydro-1H-quinolizine-2-carboxamido)-3-oxopropyl)-1,2-phenylenebis(2-methylpropanoate) hydrochloride, (2S)-isopropyl3-(3-amino-2,5-dioxo-2,3,4,5-tetrahydrobenzo[b][1,4]dioxocin-8-yl)-2-(((1-(isobutyryloxy)ethoxy)carbonyl)amino)propanoatehydrobromide, (2S)-isopropyl3-(4-amino-2,5-dioxo-2,3,4,5-tetrahydrobenzo[b][1,4]dioxocin-8-yl)-2-(((1-(isobutyryloxy)ethoxy)carbonyl)amino)propanoatehydrobromide,5-((2S)-2-(((1-(isobutyryloxy)ethoxy)carbonyl)amino-3-isopropoxy-3-oxopropyl)-2-(2-(methylamino)acetoxy)phenylbenzoate hydrochloride,4-((2S)-2-(((1-(isobutyryloxy)ethoxy)carbonyl)amino-3-isopropoxy-3-oxopropyl)-2-(2-(methylamino)acetoxy)phenylbenzoate hydrochloride,4-((2S)-3-isopropoxy-2-((((octahydroindolizin-1-yl)oxy)carbonyl)amino)-3-oxopropyl)-1,2-phenylenedibenzoate acetate, 2-(diethylamino)ethyl2-[(2,6-dichloro-3-methylphenyl)amino]benzoate.acetate,(Z)-2-(diethylaminoethyl)ethyl2-(5-fluoro-2-methyl-1-(4-methylsulfinyl)benzylidene)-1H-inden-1-yl)acetate.AcOH,2-(dimethylamino)ethyl 2-(3-phenoxyphenyl) propionate.hydrochloride,S-(2-(dimethylamino)ethyl 2-(3-phenoxyphenyl) propanethioatehydrochloride, 2-(dipropylamino)ethyl4-acetoxy-2′,4′-difluoro-[1,1′-biphenyl]-3-carboxylate hydrochloride,2-(diethylamino)ethyl 2-(4-isobutylphenyl)propionate hydrochloride,and/or 2-(diethylamino)ethyl 2-acetoxybenzoate.

In certain embodiments, the pharmaceutical composition further comprisesone or more catechol-O-methyl transferase inhibitors. In certainembodiments, the one or more catechol-O-methyl transferase inhibitorsare applied orally.

Catechol-O-methyl transferase (COMT) metabolizes L-Dopa into3-methoxy-4-hydroxy-L-phenylalanine (3-OMD) in the periphery, which doesnot easily cross the blood brain barrier (BBB). Entacapone and tolcaponeare Catechol-O-methyl transferase inhibitors and prevents COMT frommetabolizing L-DOPA into 3-methoxy-4-hydroxy-L-phenylalanine in theperiphery to avoid the undesirable effects of L-DOPA.

BERGE ET AL., 66 J. PHARM. SCI. 1-19 (1977).

For example, a pharmaceutical composition may comprise HPPs of parentdrugs or related compounds thereof, wherein the parent drugs areselected from the group consisting of Levodopa, dopamine, aspirin andother nonsteroidal anti-inflammatory drugs (NSAIDs), and anycombinations thereof plus an orally administrated catechol-O-methyltransferase inhibitor (e.g. entacapone, and tolcapone).

In certain embodiments, the pharmaceutical composition further comprisesone or more aromatic-L-amino-acid decarboxylase (DOPA Decarboxylase orDDC) inhibitors. In certain embodiments, the one or morearomatic-L-amino-acid decarboxylase inhibitors are applied orally.Aromatic-L-amino-acid decarboxylase (DOPA Decarboxylase or DDC) anenzyme important in the biosynthesis of L-DOPA to Dopamine (DA). DDCexists both outside of (body periphery) and within the confines of theblood-brain barrier. The DDC inhibitors prevent DDC conversion oflevodopa to dopamine. However, exogenously provided levadopa getsmetabolized peripherally to its active metabolite dopamine beforereaching the blood-brain barrier. Therefore, the PD brain, which isdeficient in dopamine, will not receive as much of its prodrug precursorlevodopa due to peripheral DDC breakdown. However, carbidopa and otherDDC inhibitors as a peripheral DDC inhibitor can decrease peripheral DDCconversion of levodopa before it crosses the blood-brain barrier. Inother words, carbidopa and other DDC inhibitors have no effect on brainDDC conversion of levodopa to dopamine. Ultimately, a greater proportionof the exogenously provided levodopa reaches the brain.

Examples of DDC inhibitors include, without limitation, carbidopa,benserazide, difluromethyldopa, and α-methyldopa.

In certain embodiments, a pharmaceutical composition may comprise morethan one HPPs of different parent drugs. The different parent drugs canbelong to the same or different categories of drugs that are used totreat Parkinson's disease. For example, a pharmaceutical composition maycomprise HPPs of parent drugs or related compounds thereof, wherein theparent drugs are selected from the group consisting of Levodopa,dopamine, aspirin and other nonsteroidal anti-inflammatory drugs(NSAIDs), and any combinations thereof plus one or more DDC inhibitorsselected from the group consisting of carbidopa, benserazide,difluromethyldopa, α-methyldopa, and any other aromatic-L-amino-aciddecarboxylase inhibitor.

In certain embodiments, a pharmaceutical composition comprises(S)-4-(2-amino-3-isopropoxy-3-oxopropyl)-1,2-phenylene dibenzoate and2-(diethylamino)ethyl 2-acetoxybenzoate.

In certain embodiments, a pharmaceutical composition comprises(S)-4-(2-amino-3-isopropoxy-3-oxopropyl)-1,2-phenylene dibenzoate,2-(diethylamino)ethyl 2-acetoxybenzoate, carbidopa and/or entacapone.

In certain embodiments, a pharmaceutical composition comprises(S)-4-(2-amino-3-oxo-3-(pentan-3-yloxy)propyl)-1,2-phenylenebis(2-methylpropanoate) and 2-(diethylamino)ethyl2-(4-isobutylphenyl)propionate.

In certain embodiments, a pharmaceutical composition comprises4-(2-(((1-((pyrrolidine-2-carbonyl)oxy)ethoxy)amino)ethyl)-1,2,-phenylenedibenzoate and 4-(dimethylamino)butyl 2-(3-phenoxyphenyl) propionate.

In certain embodiments, a pharmaceutical composition comprises(S)-4-(2-amino-3-oxo-3-(pentan-3-yloxy)propyl)-1,2-phenylenebis(2-methylpropanoate), 2-(diethylamino)ethyl2-(4-isobutylphenyl)propionate, carbidopa and/or entacapone.

In certain embodiments, a pharmaceutical composition comprises(S)-4-(2-amino-3-(heptan-4-yloxy)-3-oxopropyl)-1,2-phenylene dibenzoate,and 2-(dipropylamino)ethyl4-acetoxy-2′,4′-difluoro-[1,1′-biphenyl]-3-carboxylate.

In certain embodiments, a pharmaceutical composition comprises(S)-4-(2-amino-3-(heptan-4-yloxy)-3-oxopropyl)-1,2-phenylene dibenzoate,and 2-(dipropylamino)ethyl4-acetoxy-2′,4′-difluoro-[1,1′-biphenyl]-3-carboxylate, carbidopa and/orentacapone.

In certain embodiments, a pharmaceutical composition comprises atherapeutically effective amount of one or more HPPs disclosed herein.As used herein, a “therapeutically effective amount,” “therapeuticallyeffective concentration” or “therapeutically effective dose” is anamount which, as compared to a corresponding subject who has notreceived such amount, results in improved treatment, healing,prevention, or amelioration of a disease, disorder, or side effect, or adecrease in the rate of advancement of a disease or disorder.

This amount will vary depending upon a variety of factors, including butnot limited to the characteristics of the HPPs disclosed herein orpharmaceutical compositions thereof (including activity,pharmacokinetics, pharmacodynamics, and bioavailability thereof), thephysiological condition of the subject treated (including age, sex,disease type and stage, general physical condition, responsiveness to agiven dosage, and type of medication) or cells, the nature of thepharmaceutically acceptable carrier or carriers in the formulation, andthe route of administration. Further, an effective or therapeuticallyeffective amount may vary depending on whether the one or more HPPsdisclosed herein or the pharmaceutical composition thereof isadministered alone or in combination with other drug(s), othertherapy/therapies or other therapeutic method(s) or modality/modalities.One skilled in the clinical and pharmacological arts will be able todetermine an effective amount or therapeutically effective amountthrough routine experimentation, namely by monitoring a cell's orsubject's response to administration of the one or more HPPs disclosedherein or the pharmaceutical composition thereof and adjusting thedosage accordingly. A typical dosage may range from about 0.1 mg/kg toabout 100 mg/kg or more, depending on the factors mentioned above. Inother embodiments, the dosage may range from about 0.1 mg/kg to about100 mg/kg; or about 1 mg/kg to about 100 mg/kg; or about 5 mg/kg up toabout 100 mg/kg. For additional guidance, see Remington: The Science andPractice of Pharmacy, 21st Edition, Univ. of Sciences in Philadelphia(USIP), Lippincott Williams & Wilkins, Philadelphia, Pa., 2005, which ishereby incorporated by reference as if fully set forth herein foradditional guidance for determining a therapeutically effective amount.

As used herein, the term “about” refers to ±10%, ±5%, or ±1%, of thevalue following “about.”

The term “pharmaceutically acceptable carrier” as used herein means apharmaceutically-acceptable material, composition or vehicle, such as aliquid or solid filler, diluent, excipient, solvent or encapsulatingmaterial, involved in carrying or transporting an HPP from one location,body fluid, tissue, organ (interior or exterior), or portion of thebody, to another location, body fluid, tissue, organ, or portion of thebody.

Each carrier is “pharmaceutically acceptable” in the sense of beingcompatible with the other ingredients, e.g., an HPP, of the formulationand suitable for use in contact with the tissue or organ of a biologicalsubject without excessive toxicity, irritation, allergic response,immunogenicity, or other problems or complications, commensurate with areasonable benefit/risk ratio.

In certain embodiments, the pharmaceutically acceptable carrier isselected from the group consisting of alcohol, acetone, ester,cellulose, mannitol, croscarmellose sodium, vegetable oil, hydroxypropylmethylcellulose, water, and aqueous solution.

Some examples of materials which can serve aspharmaceutically-acceptable carriers include: (1) sugars, such aslactose, glucose and sucrose; (2) starches, such as corn starch andpotato starch; (3) cellulose, and its derivatives, such as sodiumcarboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4)powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients,such as cocoa butter and suppository waxes; (9) oils, such as peanutoil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil andsoybean oil; (10) glycols, such as propylene glycol; (11) polyols, suchas glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters,such as ethyl oleate and ethyl laurate; (13) agar; (14) bufferingagents, such as magnesium hydroxide and aluminum hydroxide; (15) alginicacid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer'ssolution; (19) alcohol, such as ethyl alcohol and propane alcohol; (20)phosphate buffer solutions; and (21) other non-toxic compatiblesubstances employed in pharmaceutical formulations such as acetone.

The pharmaceutical compositions may contain pharmaceutically acceptableauxiliary substances as required to approximate physiological conditionssuch as pH adjusting and buffering agents, toxicity adjusting agents andthe like, for example, sodium acetate, sodium chloride, potassiumchloride, calcium chloride, sodium lactate and the like.

In one embodiment, the pharmaceutically acceptable carrier is an aqueouscarrier, e.g. buffered saline and the like. In certain embodiments, thepharmaceutically acceptable carrier is a polar solvent, e.g. acetone andalcohol.

The concentration of HPP in these formulations can vary widely, and willbe selected primarily based on fluid volumes, viscosities, body weightand the like in accordance with the particular mode of administrationselected and the biological subject's needs. For example, theconcentration can be 0.0001% to 100%, 0.001% to 50%, 0.01% to 30%, 0.1%to 20%, 1% to 10% wt.

The compositions of the invention can be administered for prophylactic,therapeutic, and/or hygienic use. Such administration can be topical,mucosal, e.g., oral, nasal, vaginal, rectal, parenteral, transdermal,subcutaneous, intramuscular, intravenous, via inhalation, ophthalmic andother convenient routes. The pharmaceutical compositions can beadministered in a variety of unit dosage forms depending upon the methodof administration. For example, unit dosage forms suitable for oraladministration include powder, tablets, pills, capsules and lozenges andfor transdermal administration include solution, suspension and gel.

Thus, a typical pharmaceutical composition for transdermal, oral, andintravenous administrations would be about 10⁻⁸ g to about 100 g, about10⁻⁸ g to about 10⁻⁵ g, about 10⁻⁶ g to about 1 g, about 10⁻⁶ g to about100 g, about 0.001 g to about 100 g, about 0.01 g to about 10 g, orabout 0.1 g to about 1 g per subject per day. Dosages from about 0.001mg, up to about 100 g, per subject per day may be used. Actual methodsfor preparing parenterally administrable compositions will be known orapparent to those skilled in the art and are described in more detail insuch publications as Remington: The Science and Practice of Pharmacy21st ed., Lippincott Williams & Wilkins, (2005).

IV. Applications of HPPs

i) Methods for Penetrating a Biological Barrier.

Another aspect of the invention relates to a method of using acomposition of the invention in penetrating one or more biologicalbarriers in a biological subject. The method comprises a step ofadministering to a biological subject an HPP or a pharmaceuticalcomposition thereof. In certain embodiments, an HPP exhibits more thanabout 20 times or higher, 50 times or higher, >about 100 times orhigher, >about 200 time higher, >about 300 times or higher, >about 500times or higher, >about 1,000 times or higher penetration rate throughone or more biological barriers than its parent drug.

The term “biological barrier” as used herein refers to a biologicallayer that separates an environment into different spatial areas orcompartments, which separation is capable of modulating (e.g.restricting, limiting, enhancing or taking no action in) the passingthrough, penetrating or translocation of substance or matter from onecompartment/area to another. The different spatial areas or compartmentsas referred to herein may have the same or different chemical orbiological environment(s). The biological layer as referred hereinincludes, but is not limited to, a biological membrane, a cell layer, abiological structure, an inner surface of subjects, organisms, organs orbody cavities, an external surface of subjects, organisms, organs orbody cavities, or any combination or plurality thereof.

Examples of a biological membrane include a lipid bilayer structure,eukaryotic cell membrane, prokaryotic cell membrane, and intracellularmembrane (e.g., nucleus or organelle membrane, such as membrane orenvelope of Golgi apparatus, rough and smooth endoplasmic reticulum(ER), ribosomes, vacuoles, vesicles, liposomes, mitochondria, lysosome,nucleus, chloroplasts, plastids, peroxisomes or microbodies).

The lipid bilayer referred to herein is a double layer of lipid-classmolecules, including, but not limited to, phospholipids and cholesterol.In a particular embodiment, lipids for bilayer are amphiphilic moleculesconsisting of polar head groups and non-polar fatty acid tails. Thebilayer is composed of two layers of lipids arranged so that theirhydrocarbon tails face one another to form an oily core held together bythe hydrophobic effect, while their charged heads face the aqueoussolutions on either side of the membrane. In another particularembodiment, the lipid bilayer may contain one or more embedded proteinand/or sugar molecule(s).

Examples of a cell layer include a lining of eukaryotic cells (e.g.,epithelium, lamina propria and smooth muscle or muscularis mucosa (ingastrointestinal tract)), a lining of prokaryotic cells (e.g., surfacelayer or S-layer which refers to a two dimensional structuremonomolecular layer composed of identical proteins or glycoproteins,specifically, an S-layer refers to a part of a cell envelope commonlyfound in bacteria and archaea), a biofilm (a structured community ofmicroorganisms encapsulated within a self-developed polymeric matrix andadherent to a living or inert surface), and a plant cell layer (e.g.,epidermis). The cells may be normal cells or pathological cells (e.g.disease cells, cancer cells).

Examples of biological structures include structures sealed by tight oroccluding junctions that provide a barrier to the entry of toxins,bacteria and viruses, e.g. the blood milk barrier and the blood brainbarrier (BBB). In particular, BBB is composed of an impermeable class ofendothelium, which presents both a physical barrier through tightjunctions adjoining neighboring endothelial cells and a transportbarrier comprised of efflux transporters. The biological structure mayalso include a mixture of cells, proteins and sugars (e.g. blood clots).

Examples of the inner surface of subjects, organisms, organs or bodycavities include buccal mucosa, esophageal mucosa, gastric mucosa,intestinal mucosa, olfactory mucosa, oral mucosa, bronchial mucosa,uterine mucosa and endometrium (the mucosa of the uterus, inner layer ofthe wall of a pollen grain or the inner wall layer of a spore), or acombination or plurality thereof.

Examples of the external surface of subjects, organisms, organs or bodycavities include capillaries (e.g. capillaries in the heart tissue),mucous membranes that are continuous with skin (e.g. such as at thenostrils, the lips, the ears, the genital area, and the anus), outersurface of an organ (e.g. liver, lung, stomach, brain, kidney, heart,ear, eye, nose, mouth, tongue, colon, pancreas, gallbladder, duodenum,rectum stomach, colonrectum, intestine, vein, respiratory system,vascular, anorectum and pruritus ani), skin, cuticle (e.g. dead layersof epidermal cells or keratinocytes or superficial layer of overlappingcells covering the hair shaft of an animal, a multi-layered structureoutside the epidermis of many invertebrates, plant cuticles or polymerscutin and/or cutan), external layer of the wall of a pollen grain or theexternal wall layer of a spore), and a combination or plurality thereof.

In addition, a biological barrier further includes a sugar layer, aprotein layer or any other biological layer, or a combination orplurality thereof. For example, skin is a biological barrier that has aplurality of biological layers. A skin comprises an epidermis layer(outer surface), a demis layer and a subcutaneous layer. The epidermislayer contains several layers including a basal cell layer, a spinouscell layer, a granular cell layer, and a stratum corneum. The cells inthe epidermis are called keratinocytes. The stratum corneum (“hornylayer”) is the outmost layer of the epidermis, wherein cells here areflat and scale-like (“squamous”) in shape. These cells contain a lot ofkeratin and are arranged in overlapping layers that impart a tough andoilproof and waterproof character to the skin's surface.

ii) Methods for Treating Parkinson's Disease in a Biological Subject

Another aspect of the invention relates to a method of using acomposition of the invention, or a pharmaceutical composition thereof intreating a condition in a biological subject. The method comprisesadministrating the pharmaceutical composition to the biological subject.

The term “treating” as used herein means curing, alleviating,inhibiting, or preventing. The term “treat” as used herein means cure,alleviate, inhibit, or prevent. The term “treatment” as used hereinmeans cure, alleviation, inhibition or prevention.

The term “biological subject,” or “subject” as used herein means anorgan, a group of organs that work together to perform a certain task,an organism, or a group of organisms. The term “organism” as used hereinmeans an assembly of molecules that function as a more or less stablewhole and has the properties of life, such as animal, plant, fungus, ormicro-organism.

The term “animal” as used herein means a eukaryotic organismcharacterized by voluntary movement. Examples of animals include,without limitation, vertebrata (e.g. human, mammals, birds, reptiles,amphibians, fishes, marsipobranchiata and leptocardia), tunicata (e.g.thaliacea, appendicularia, sorberacea and ascidioidea), articulata (e.g.insecta, myriapoda, malacapoda, arachnida, pycnogonida, merostomata,crustacea and annelida), gehyrea (anarthropoda), and helminthes (e.g.rotifera).

The conditions the method can treat include conditions that can betreated by the parent drug of the HPP are Parkinson's disease andrelated conditions.

iii). Methods of Using One or More HPPs or a Pharmaceutical CompositionsThereof in Treatments of Parkinson's Disease and Related Conditions.

Another aspect of the invention relates to a method of using one or moreHPPs or a pharmaceutical composition thereof in treating Parkinson'sdisease and/or related conditions in a biological subject or subject byadministrating the one or more HPPs or a pharmaceutical compositionthereof to the biological subject or subject.

In certain embodiments, a method of treating a Parkinson's disease andrelated conditions in a subject comprises administering a therapeuticeffective amount of an HPP, or a pharmaceutical composition thereof tothe subject.

An HPP or a pharmaceutical composition thereof can be administered to abiological subject by any administration route known in the art,including without limitation, oral, enteral, buccal, nasal, topical,rectal, vaginal, aerosol, transmucosal, epidermal, transdermal, dermal,ophthalmic, pulmonary, subcutaneous, and/or parenteral administration.The pharmaceutical compositions can be administered in a variety of unitdosage forms depending upon the method of administration.

The preferred administered methods are transdermal, dermal, topicaladministration for a compound.

In certain embodiments, the method further comprises orallyadministering to the subject a therapeutically effective amount of oneor more COMT inhibitors and/or one or more DDC inhibitors describedsupra.

When a plurality of drugs (e.g. HPPs, COMT inhibitors and DDCinhibitors) are applied to a subject, the plurality of HPPs and/or otherdrugs can be applied at substantially the same time or at differenttimes. The plurality of drugs may be mixed together before theadministration to the subject, or administered to the subjectseparately. In certain embodiments, some of the plurality of drugs aremixed before administration, while others are applied separately. Eachdrug may be applied in any possible order, and any possible manner.

As used herein, a pharmaceutical composition may comprise multiplecomponents that are applied to the subject in different administrationroute. For example, a pharmaceutical composition comprising one or moreHPPs and one or more COMT inhibitors and/or one or more DDC inhibitorsdescribed supra, the one or more HPPs may be applied transdermally,topically, or dermally, while the one or more COMT inhibitors and/or oneor more DDC inhibitors described supra may be applied orally. Themultiple components may be applied in any order and any combinationpossible.

An HPP or a pharmaceutical composition thereof can be given to a subjectin the form of formulations or preparations suitable for eachadministration route. The formulations useful in the methods of theinvention include one or more HPPs, one or more pharmaceuticallyacceptable carriers therefor, and optionally other therapeuticingredients. The formulations may conveniently be presented in unitdosage form and may be prepared by any methods well known in the art ofpharmacy. The amount of active ingredient which can be combined with acarrier material to produce a single dosage form will vary dependingupon the subject being treated and the particular mode ofadministration. The amount of an HPP which can be combined with acarrier material to produce a pharmaceutically effective dose willgenerally be that amount of an HPP which produces a therapeutic effect.Generally, out of one hundred percent, this amount will range from about1 percent to about ninety-nine percent of the HPP, preferably from about1 percent to about 20 percent.

Methods of preparing these formulations or compositions include the stepof bringing into association an HPP with one or more pharmaceuticallyacceptable carriers and, optionally, one or more accessory ingredients.In general, the formulations are prepared by uniformly and intimatelybringing into association an HPP with liquid carriers, or finely dividedsolid carriers, or both, and then, if necessary, shaping the product.

Formulations suitable for oral administration may be in the form ofcapsules, cachets, pills, tablets, lozenges (using a flavored basis,usually sucrose and acacia or tragacanth), powders, granules, or as asolution or a suspension in an aqueous or non-aqueous liquid, or as anoil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup,or as pastilles (using an inert base, such as gelatin and glycerin, orsucrose and acacia) and/or as mouth washes and the like, each containinga predetermined amount of an HPP as an active ingredient. A compound mayalso be administered as a bolus, electuary, or paste.

In solid dosage forms for oral administration (e. g., capsules, tablets,pills, dragees, powders, granules and the like), the HPP is mixed withone or more pharmaceutically-acceptable carriers, such as sodium citrateor dicalcium phosphate, and/or any of the following: (1) fillers orextenders, such as starches, lactose, sucrose, glucose, mannitol, and/orsilicic acid; (2) binders, such as, for example, carboxymethylcellulose,alginates, gelatin, polyvinyl pyrrolidone, sucrose and/or acacia; (3)humectants, such as glycerol; (4) disintegrating agents, such asagar-agar, calcium carbonate, potato or tapioca starch, alginic acid,certain silicates, and sodium carbonate, (5) solution retarding agents,such as paraffin, (6) absorption accelerators, such as quaternaryammonium compounds; (7) wetting agents, such as, for example, acetylalcohol and glycerol monostearate; (8) absorbents, such as kaolin andbentonite clay; (9) lubricants, such a talc, calcium stearate, magnesiumstearate, solid polyethylene glycols, sodium lauryl sulfate, andmixtures thereof; and (10) coloring agents. In the case of capsules,tablets and pills, the pharmaceutical compositions may also comprisebuffering agents. Solid compositions of a similar type may also beemployed as fillers in soft and hard-filled gelatin capsules using suchexcipients as lactose or milk sugars, as well as high molecular weightpolyethylene glycols and the like.

A tablet may be made by compression or molding, optionally with one ormore accessory ingredients. Compressed tablets may be prepared usingbinder (for example, gelatin or hydroxypropylmethyl cellulose),lubricant, inert diluent, preservative, disintegrant (for example,sodium starch glycolate or cross-linked sodium carboxymethyl cellulose),surface-active or dispersing agent. Molded tablets may be made bymolding in a suitable machine a mixture of the powdered HPPs orpeptidomimetic moistened with an inert liquid diluent. Tablets, andother solid dosage forms, such as dragees, capsules, pills and granules,may optionally be scored or prepared with coatings and shells, such asenteric coatings and other coatings well known in thepharmaceutical-formulating art. They may also be formulated so as toprovide slow or controlled release of an HPP therein using, for example,hydroxypropylmethyl cellulose in varying proportions to provide thedesired release profile, other polymer matrices, liposomes and/ormicrospheres. They may be sterilized by, for example, filtration througha bacteria-retaining filter, or by incorporating sterilizing agents inthe form of sterile solid compositions which can be dissolved in sterilewater, or some other sterile injectable medium immediately before use.These compositions may also optionally contain pacifying agents and maybe of a composition that they release the HPP(s) only, orpreferentially, in a certain portion of the gastrointestinal tract,optionally, in a delayed manner. Examples of embedding compositionswhich can be used include polymeric substances and waxes. The HPP canalso be in micro-encapsulated form, if appropriate, with one or more ofthe above-described excipients.

Liquid dosage forms for oral administration include pharmaceuticallyacceptable emulsions, microemulsions, solutions, suspensions, syrups andelixirs. In addition to the HPP, the liquid dosage forms may containinert diluents commonly used in the art, such as, for example, water orother solvents, solubilizing agents and emulsifiers, such as ethylalcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzylalcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils(in particular, cottonseed, groundnut, corn, germ, olive, castor andsesame oils), glycerol, tetrahydrofuryl alcohol, polyethylene glycolsand fatty acid esters of sorbitan, and mixtures thereof. Besides inertdiluents, the oral compositions can also include adjuvants such aswetting agents, emulsifying and suspending agents, sweetening,flavoring, coloring, perfuming and preservative agents.

Suspensions, in addition to the HPP, may contain suspending agents as,for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitoland sorbitan esters, microcrystalline cellulose, aluminum metahydroxide,bentonite, agar-agar and tragacanth, and mixtures thereof.

Formulations for rectal or vaginal administration may be presented as asuppository, which may be prepared by mixing one or more HPPs with oneor more suitable nonirritating excipients or carriers comprising, forexample, cocoa butter, polyethylene glycol, a suppository wax or asalicylate, and which is solid at room temperature, but liquid at bodytemperature and, therefore, will melt in the rectum or vaginal cavityand release the active agent. Formulations which are suitable forvaginal administration also include pessaries, tampons, creams, gels,pastes, foams or spray formulations containing such carriers as areknown in the art to be appropriate.

Formulations for the topical or transdermal or epidermal or dermaladministration of an HPP composition include powders, sprays, ointments,pastes, creams, lotions, gels, solutions, patches and inhalants. Theactive component may be mixed under sterile conditions with apharmaceutically acceptable carrier, and with any preservatives,buffers, or propellants which may be required. The ointments, pastes,creams and gels may contain, in addition to the HPP composition,excipients, such as animal and vegetable fats, oils, waxes, paraffins,starch, tragacanth, cellulose derivatives, polyethylene glycols,silicones, bentonites, silicic acid, talc and zinc oxide, or mixturesthereof. Powders and sprays can contain, in addition to the HPPcomposition, excipients such as lactose, talc, silicic acid, aluminumhydroxide, calcium silicates and polyamide powder, or mixtures of thesesubstances. Sprays can additionally contain customary propellants, suchas chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons,such as butane and propane. The best formulations for the topical ortransdermal administration are pure water, solution, aqueous solution,ethanol and water solution, and isopropanol and water solution.

An HPP or a pharmaceutical composition thereof can be alternativelyadministered by aerosol. This can be accomplished by preparing anaqueous aerosol, liposomal preparation or solid particles containing theHPPs. A nonaqueous (e. g., fluorocarbon propellant) suspension could beused. Sonic nebulizers can also be used. An aqueous aerosol is made byformulating an aqueous solution or suspension of the agent together withconventional pharmaceutically acceptable carriers and stabilizers. Thecarriers and stabilizers vary with the requirements of the particularcompound, but typically include nonionic surfactants (Tweens, Pluronics,or polyethylene glycol), innocuous proteins like serum albumin, sorbitanesters, oleic acid, lecithin, amino acids such as glycine, buffers,salts, sugars or sugar alcohols. Aerosols generally are prepared fromisotonic solutions.

Transdermal patches can also be used to deliver HPP compositions to atarget site. Such formulations can be made by dissolving or dispersingthe agent in the proper medium. Absorption enhancers can also be used toincrease the flux of the peptidomimetic across the skin. The rate ofsuch flux can be controlled by either providing a rate controllingmembrane or dispersing the peptidomimetic in a polymer matrix or gel.

Ophthalmic formulations, eye ointments, powders, solutions and the like,are also contemplated as being within the scope of this invention.

Formulations suitable for parenteral administration comprise an HPP incombination with one or more pharmaceutically-acceptable sterileisotonic aqueous or nonaqueous solutions, dispersions, suspensions oremulsions, or sterile powders which may be reconstituted into sterileinjectable solutions or dispersions just prior to use, which may containantioxidants, buffers, bacteriostats, solutes which render theformulation isotonic with the blood of the intended recipient orsuspending or thickening agents.

Examples of suitable aqueous and nonaqueous carriers which may beemployed in the formulations suitable for parenteral administrationinclude water, ethanol, polyols (e. g., such as glycerol, propyleneglycol, polyethylene glycol, and the like), and suitable mixturesthereof, vegetable oils, such as olive oil, and injectable organicesters, such as ethyl oleate. Proper fluidity can be maintained, forexample, by the use of coating materials, such as lecithin, by themaintenance of the required particle size in the case of dispersions,and by the use of surfactants.

Formulations suitable for parenteral administration may also containadjuvants such as preservatives, wetting agents, emulsifying agents anddispersing agents. Prevention of the action of microorganisms may beensured by the inclusion of various antibacterial and antifungal agents,for example, paraben, chlorobutanol, phenol sorbic acid, and the like.It may also be desirable to include isotonic agents, such as sugars,sodium chloride, and the like into the compositions. In addition,prolonged absorption of the injectable pharmaceutical form may bebrought about by the inclusion of agents which delay absorption such asaluminum monostearate and gelatin.

Injectable depot forms are made by forming microencapsule matrices of anHPP or in biodegradable polymers such as polylactide-polyglycolide.Depending on the ratio of the HPP to polymer, and the nature of theparticular polymer employed, the rate of drug release can be controlled.Examples of other biodegradable polymers include poly (orthoesters) andpoly (anhydrides). Depot injectable formulations are also prepared byentrapping the HPP in liposomes or microemulsions which are compatiblewith body tissue.

In certain embodiments, an HPP or a pharmaceutical composition thereofis delivered to an action site in a therapeutically effective dose. Asis known in the art of pharmacology, the precise amount of thepharmaceutically effective dose of an HPP that will yield the mosteffective results in terms of efficacy of treatment in a given patientwill depend upon, for example, the activity, the particular nature,pharmacokinetics, pharmacodynamics, and bioavailability of a particularHPP, physiological condition of the subject (including race, age, sex,weight, diet, disease type and stage, general physical condition,responsiveness to a given dosage and type of medication), the nature ofpharmaceutically acceptable carriers in a formulation, the route andfrequency of administration being used, and the severity or propensityof the condition that is to be treated. However, the above guidelinescan be used as the basis for fine-tuning the treatment, e. g.,determining the optimum dose of administration, which will require nomore than routine experimentation consisting of monitoring the subjectand adjusting the dosage. Remington: The Science and Practice ofPharmacy (Gennaro ed. 20.sup.th edition, Williams & Wilkins PA, USA)(2000).

IV. Advantages

Levodopa may cause nausea, vomiting, gastrointestinal bleeding,dyskinesia at peak dose, and end-of-dose deterioration of function.These problems may be solved by transdermally administration of the highpenetration compositions of L-Dopa and dopamine which can deliver aminimal therapeutically effective amount constantly to avoidgastrointestinal bleeding, dyskinesia at peak dose, and end-of-dosedeterioration of function.

Recently, increasing evidence from human and animal studies hassuggested that neuroinflammation is an important contributor to theneuronal loss in PD. Moreover, the pro-inflammatory agentlipopolysaccharide itself can directly initiate degeneration ofdopamine-containing neurons or combine with other environmentalfactor(s), such as the pesticide rotenone, to exacerbate suchneurodegeneration. These effects provide strong support for theinvolvement of inflammation in the pathogenesis of PD. Although dopaminereplacement can alleviate symptoms of the disorder, there is no proventherapy to halt the underlying progressive degeneration ofdopamine-containing neurons. The transdermally administration of thehigh penetration compositions of L-Dopa and/or dopamine with the highpenetration compositions of aspirin and/or other NSAID may not onlyalleviate symptoms of Parkinson's disease, but also halt the underlyingprogressive degeneration of dopamine containing neurons.

When the high penetration compositions of L-Dopa and/or dopamine, andaspirin and/or other NSAID are administered transdermally, Carbidopa,benserazide, difluromethyldopa, α-methyldopa, and/or other DDCinhibitors and/or and/or entacapone, andtolcapone, and/or other COMTinhibitors are administrated orally can help a greater proportion of theexogenously provided levodopa or dopamine reaches the brain to reducethe L-dopa or dopamine side effects.

V. Examples

The following examples are provided to better illustrate the claimedinvention and are not to be interpreted in any way as limiting the scopeof the invention. All specific compositions, materials, and methodsdescribed below, in whole or in part, fall within the scope of theinvention. These specific compositions, materials, and methods are notintended to limit the invention, but merely to illustrate specificembodiments falling within the scope of the invention. One skilled inthe art may develop equivalent compositions, materials, and methodswithout the exercise of inventive capacity and without departing fromthe scope of the invention. It will be understood that many variationscan be made in the procedures herein described while still remainingwithin the bounds of the invention. It is the intention of the inventorsthat such variations are included within the scope of the invention.Furthermore, all references cited herein are incorporated by referencein their entireties, as if fully set forth herein.

Example 1 Preparation of4-(2-(2-(methylamino)acetamido)ethyl)-1,2,-phenylene dibenzoatehydrochloride

In this example, a dopamine.HCl solution was prepared by addingdopamine.HCl (19 g) into acetone (200 mL). Water (200 mL) and sodiumbicarbonate (R0090, 50 g) were added into the dopamine.HCl solution toprovide a 1st mixture. Then Boc-N-methylglycine N-hydroxysuccinimideester (Boc-Sar-OSu, 29 g) was added to the 1st mixture to provide a 2ndmixture. The 2nd mixture was stirred at RT overnight, and added ethylacetate (R0061, 500 mL) for extraction. The organic layer was separated,and washed with water (2×100 mL), 20% citric acid (2×200 mL), and water(3×100 mL), respectively, and dried over sodium sulfate to provide a 3rdsolution. Then the sodium sulfate was filtered and washed with ethylacetate to provide a 4th mixture. The 3rd solution and the 4th mixturewere combined, and added first with pyridine (R0081, 30 mL), and thenwith benzoyl chloride (R0488, 30 g) drop by drop to provide a 5thmixture. The 5th mixture was stirred at RT for 2 hours, then washed withwater (2×100 mL), 5% sodium bucarbonate (2×100 mL), water (100 mL), 20%citric acid (2×200 mL), and water (3×100 mL), respectively, and driedover sodium sulfate to provide a 6th solution. The sodium sulfate wasfiltered and washed with ethyl acetate to provide a 7th solution. The6th solution and the 7th solution were combined and concentrated toabout 200 mL (an 8th solution). Into the 8th solution was added anisole(20 g), and then bubbled HCl gas (20 g) to form precipitation. Theprecipitated solid was collected and washed with ethyl acetate to yield4-(2-(2-(methylamino)acetamido)ethyl)-1,2-phenylene dibenzoatehydrochloride salt.

Example 2 Preparation of4-(2-(2-(methylamino)acetamido)ethyl)-1,2,-phenylene dibenzoate aceticacid

In this example, 4-(2-(2-(methylamino)acetamido)ethyl)-1,2,-phenylenedibenzoate hydrochloride (23 g) was dissolved in isopropanol (300 mL) toprovide a 1st mixture. Sodium acetate (4 g) was added into the 1stmixture to provide a 2nd mixture. The 2nd mixture was stirred at RT for2 hours, and filtered to remove solid therein. The filtered solution wasevaporated to dryness to yield4-(2-(2-(methylamino)acetamido)ethyl)-1,2,-phenylene dibenzoate aceticacid salt.

Example 3 Preparation of4-(6-methyl-4,8-dioxo-5,7-dioxa-2,9-diazadecan-11-yl)-1,2,-phenylenedibenzoate hydrobromide

In this example, a dopamine.HCl solution was prepared by addingdopamine.HCl (19 g) into acetone (200 mL). Water (200 mL) and sodiumbicarbonate (R0090, 50 g) were added into the dopamine.HCl solution toprovide a 1st mixture. Then di-tert-butyl dicarbonate (22 g) was addedto the 1st mixture to provide a 2nd mixture. The 2nd mixture was stirredat RT overnight, and added ethyl acetate (R0061, 500 mL) for extraction.The organic layer was separated, and washed with water (2×100 mL), 20%citric acid (2×200 mL), and water (3×100 mL), respectively, and driedover sodium sulfate to provide a 3rd solution. Then the sodium sulfatewas filtered and washed with ethyl acetate to provide a 4th mixture. The3rd solution and the 4th mixture were combined, and added first withpyridine (R0081, 30 mL), and then with benzoyl chloride (R0488, 30 g)drop by drop to provide a 5th mixture. The 5th mixture was stirred at RTfor 2 hours, then washed with water (2×100 mL), 5% sodium bucarbonate(2×100 mL), water (100 mL), 20% citric acid (2×200 mL), and water (3×100mL), respectively, and dried over sodium sulfate to provide a 6thsolution. The sodium sulfate was filtered and washed with ethyl acetateto provide a 7th solution. The 6th solution and the 7th solution werecombined and concentrated to about 200 mL (an 8th solution). Into the8th solution was added anisole (20 g), and then bubbled HCl gas (20 g)to form precipitation. The precipitated solid was collected and washedwith ethyl acetate, and then suspended in DCM (200 mL) to provide a 9thmixture. Sodium bicarbonate (20 g) and tetrabutylammonium hydrogensulfate (11 g) were added into the 9th mixture to provide a 10thmixture. Then 1-chloroethyl chloroformate (16 g) was added into the 10thmixture to provide an 11th mixture. The 11th mixture was stirred at RTovernight. Then the organic layer of the 11th mixture was collected andwashed with water (3×200 mL), and dried over anhydrous sodium sulfate toprovide a 12th solution. The sodium sulfate was removed by filtrationand washed with DCM to provide a 13th solution. The 12th solution andthe 13th solution were combined and evaporated to dryness. The residuewas dissolved in acetonitrile (200 mL) to provide a 14th mixture.Boc-sarcosine (36 g) was added into the 14th mixture to provide a 15thmixture. A mixture of [diisopropylethylamine (34 mL) and Boc-sarcosine(36 g) was prepared first and then added into the 15th mixture toprovide a 16th mixture. The 16th mixture was stirred at 55° C. for 48hours, and added ethyl acetate (500 mL) with stirring. The obtainedorganic layer was collected and washed with 5% sodium bicarbonate (3×100mL) and water (3×100 mL), respectively, and dried over anhydrous sodiumsulfate to provide a 17th solution. The sodium sulfate was filtered andwashed with ethyl acetate to provide a 18th solution. The 17th solutionand the 18th solution were combined and concentrated to 300 mL. Into theconcentrated solution was added anisole (20 g) and then bubbled HBr gas(30 g) to provide precipitation. The precipitated solid was collectedand washed with ethyl acetate to yield4-(6-methyl-4,8-dioxo-5,7-dioxa-2,9-diazadecan-11-yl)-1,2,-phenylenedibenzoate hydrobromide.

Example 4 Preparation of4-(2-(2-amino-3-phenylpropanamido)ethyl)-1,2,-phenylene dibenzoatehydrochloride

In this example, a dopamine.HCl solution was prepared by addingdopamine.HCl (19 g) into acetone (200 mL). Water (200 mL) and sodiumbicarbonate (R0090, 50 g) were added into the dopamine.HCl solution toprovide a 1st mixture. Then boc-phenylalanine N-hydroxysuccinimide ester(Boc-Phe-OSu, 36 g) was added to the 1st mixture to provide a 2ndmixture. The 2nd mixture was stirred at RT overnight, and added ethylacetate (R0061, 500 mL) for extraction. The organic layer was separated,and washed with water (2×100 mL), 20% citric acid (2×200 mL), and water(3×100 mL), respectively, and dried over sodium sulfate to provide a 3rdsolution. Then the sodium sulfate was filtered and washed with ethylacetate to provide a 4th mixture. The 3rd solution and the 4th mixturewere combined, and added first with pyridine (R0081, 30 mL), and thenwith benzoyl chloride (R0488, 30 g) drop by drop to provide a 5thmixture. The 5th mixture was stirred at RT for 2 hours, then washed withwater (2×100 mL), 5% sodium bucarbonate (2×100 mL), water (100 mL), 20%citric acid (2×200 mL), and water (3×100 mL), respectively, and driedover sodium sulfate to provide a 6th solution. The sodium sulfate wasfiltered and washed with ethyl acetate to provide a 7th solution. The6th solution and the 7th solution were combined and concentrated toabout 200 mL (an 8th solution). Into the 8th solution was added anisole(20 g), and then bubbled HCl gas (20 g) to form precipitation. Theprecipitated solid was collected and washed with ethyl acetate to yield4-(2-(2-amino-3-phenylpropanamido)ethyl)-1,2,-phenylene dibenzoatehydrochloride.

Example 5 Preparation of4-(2-(((1-((pyrrolidine-2-carbonyl)oxy)ethoxy)amino)ethyl)-1,2,-phenylenedibenzoate hydrochloride

In this example, a dopamine.HCl solution was prepared by addingdopamine.HCl (19 g) into acetone (200 mL). Water (200 mL) and sodiumbicarbonate (R0090, 50 g) were added into the dopamine.HCl solution toprovide a 1st mixture. Then di-tert-butyl dicarbonate (22 g) was addedto the 1st mixture to provide a 2nd mixture. The 2nd mixture was stirredat RT overnight, and added ethyl acetate (R0061, 500 mL) for extraction.The organic layer was separated, and washed with water (2×100 mL), 20%citric acid (2×200 mL), and water (3×100 mL), respectively, and driedover sodium sulfate to provide a 3rd solution. Then the sodium sulfatewas filtered and washed with ethyl acetate to provide a 4th mixture. The3rd solution and the 4th mixture were combined, and added first withpyridine (R0081, 30 mL), and then with benzoyl chloride (R0488, 30 g)drop by drop to provide a 5th mixture. The 5th mixture was stirred at RTfor 2 hours, then washed with water (2×100 mL), 5% sodium bucarbonate(2×100 mL), water (100 mL), 20% citric acid (2×200 mL), and water (3×100mL), respectively, and dried over sodium sulfate to provide a 6thsolution. The sodium sulfate was filtered and washed with ethyl acetateto provide a 7th solution. The 6th solution and the 7th solution werecombined and concentrated to about 200 mL (an 8th solution). Into the8th solution was added anisole (20 g), and then bubbled HCl gas (20 g)to form precipitation. The precipitated solid was collected and washedwith ethyl acetate, and then suspended in DCM (200 mL) to provide a 9thmixture. Sodium bicarbonate (20 g) and tetrabutylammonium hydrogensulfate (11 g) were added into the 9th mixture to provide a 10thmixture. Then 1-chloroethyl chloroformate (16 g) was added into the 10thmixture to provide an 11th mixture. The 11th mixture was stirred at RTovernight. Then the organic layer of the 11th mixture was collected andwashed with water (3×200 mL), and dried over anhydrous sodium sulfate toprovide a 12th solution. The sodium sulfate was removed by filtrationand washed with DCM to provide a 13th solution. The 12th solution andthe 13th solution were combined and evaporated to dryness. The residuewas dissolved in acetonitrile (200 mL) to provide a 14th mixture.Boc-sarcosine (36 g) was added into the 14th mixture to provide a 15thmixture. A mixture of [diisopropylethylamine (34 mL) and Boc-sarcosine(36 g) was prepared first and then added into the 15th mixture toprovide a 16th mixture. The 16th mixture was stirred at 55° C. for 48hours, and added ethyl acetate (500 mL) with stirring. The obtainedorganic layer was collected and washed with 5% sodium bicarbonate (3×100mL) and water (3×100 mL), respectively, and dried over anhydrous sodiumsulfate to provide a 17th solution. The sodium sulfate was filtered andwashed with ethyl acetate to provide a 18th solution. The 17th solutionand the 18th solution were combined and concentrated to 300 mL. Into theconcentrated solution was added anisole (20 g) and then bubbled HCl gas(30 g) to provide precipitation. The precipitated solid was collectedand washed with ethyl acetate to yield4-(2-(((1-((pyrrolidine-2-carbonyl)oxy)ethoxy)amino)ethyl)-1,2,-phenylenedibenzoate hydrochloride.

Example 6 Preparation of4-(2-piperidine-4-carboxamido)ethyl)-1,2,-phenylenebis(2-ethylbutanoate) hydrochloride

In this example, a dopamine.HCl solution was prepared by addingdopamine.HCl (19 g) into acetone (200 mL). Water (200 mL) and sodiumbicarbonate (R0090, 50 g) were added into the dopamine.HCl solution toprovide a 1st mixture. Then Boc-piperidine-4-carboxylic acidN-hydroxysuccinimide ester (Boc-Inp-OSu, 33 g) was added to the 1stmixture to provide a 2nd mixture. The 2nd mixture was stirred at RTovernight, and added ethyl acetate (R0061, 500 mL) for extraction. Theorganic layer was separated, and washed with water (2×100 mL), 20%citric acid (2×200 mL), and water (3×100 mL), respectively, and driedover sodium sulfate to provide a 3rd solution. Then the sodium sulfatewas filtered and washed with ethyl acetate to provide a 4th mixture. The3rd solution and the 4th mixture were combined, and added first withpyridine (R0081, 30 mL), and then with 2-ethylbutyryl chloride (28 g)drop by drop to provide a 5th mixture. The 5th mixture was stirred at RTfor 2 hours, then washed with water (2×100 mL), 5% sodium bucarbonate(2×100 mL), water (100 mL), 20% citric acid (2×200 mL), and water (3×100mL), respectively, and dried over sodium sulfate to provide a 6thsolution. The sodium sulfate was filtered and washed with ethyl acetateto provide a 7th solution. The 6th solution and the 7th solution werecombined and concentrated to about 200 mL (an 8th solution). Into the8th solution was added anisole (20 g), and then bubbled HCl gas (20 g)to form precipitation. The precipitated solid was collected and washedwith ethyl acetate to yield4-(2-piperidine-4-carboxamido)ethyl)-1,2,-phenylenebis(2-ethylbutanoate) hydrochloride.

Example 7 Preparation of4-(2-((((-octahydro-1H-quinolizin-3-yl)oxy)carbonyl)amino)ethyl)-1,2,-phenylenebis(2-ethylbutanoate)acetate

In this example, a dopamine.HCl solution was prepared by addingdopamine.HCl (19 g) into acetone (200 mL). Water (200 mL) and sodiumbicarbonate (R0090, 50 g) were added into the dopamine.HCl solution toprovide a 1st mixture. Then di-tert-butyl dicarbonate (22 g) was addedto the 1st mixture to provide a 2nd mixture. The 2nd mixture was stirredat RT overnight, and added ethyl acetate (R0061, 500 mL) for extraction.The organic layer was separated, and washed with water (2×100 mL), 20%citric acid (2×200 mL), and water (3×100 mL), respectively, and driedover sodium sulfate to provide a 3rd solution. Then the sodium sulfatewas filtered and washed with ethyl acetate to provide a 4th mixture. The3rd solution and the 4th mixture were combined, and added first withpyridine (R0081, 30 mL), and then with 2-ethylbutyryl chloride (28 g)drop by drop to provide a 5th mixture. The 5th mixture was stirred at RTfor 2 hours, then washed with water (2×100 mL), 5% sodium bucarbonate(2×100 mL), water (100 mL), 20% citric acid (2×200 mL), and water (3×100mL), respectively, and dried over sodium sulfate to provide a 6thsolution. The sodium sulfate was filtered and washed with ethyl acetateto provide a 7th solution. The 6th solution and the 7th solution werecombined and concentrated to about 200 mL (an 8th solution). Into the8th solution was added anisole (20 g), and then bubbled HCl gas (20 g)to form precipitation. The precipitated solid was collected and washedwith ethyl acetate to yield4-(2-((((-octahydro-1H-quinolizin-3-yl)oxy)carbonyl)amino)ethyl)-1,2,-phenylenebis(2-ethylbutanoate) hydrochloride salt.

The solid was suspended in ethyl acetate (200 mL) to provide a 9thmixture. Into the 9th mixture was first added triethylamine (25 mL), andthen drop by drop with octahydro-1H-quinolizin-3-yl carbonochloridehydrochloride (22 g) in ethyl acetate (50 mL) to provide a 10th mixture.The 10th mixture was stirred for 2 hours at RT, washed with 5% sodiumbicarbonate and water to provide a 11th mixture. Acetic acid (6 g) wasadded into the 11th mixture, followed by addition of hexane (200 mL) toprovide precipitation. The precipitated solid was collected byfiltration and washed with ethyl acetate/hexanes to yield4-(2-((((-octahydro-1H-quinolizin-3-yl)oxy)carbonyl)amino)ethyl)-1,2,-phenylenebis(2-ethylbutanoate)acetate.

Example 8 Preparation of1-(((2-(4-amino-2,5-dioxo-2,3,4,5-tetrahydrobenzo[b][1,4]dioxocin-8-yl)ethyl)carbamoyl)oxy)ethylisobutyrate hydrofluoride and1-(((2-(3-amino-2,5-dioxo-2,3,4,5-tetrahydrobenzo[b][1,4]dioxocin-8-yl)ethyl)carbamoyl)oxy)ethylisobutyrate hydrofluoride

In this example, a dopamine.HCl solution was prepared by addingdopamine.HCl (19 g) into acetone (200 mL). Water (200 mL) and sodiumbicarbonate (R0090, 50 g) were added into the dopamine.HCl solution toprovide a 1st mixture. Then Nα-(benzyloxycarbonyloxy)succinimide (25 g)was added to the 1st mixture to provide a 2nd mixture. The 2nd mixturewas stirred at RT overnight, and added ethyl acetate (R0061, 500 mL) forextraction. The organic layer was separated, and washed with water(2×100 mL), 20% citric acid (2×200 mL), and water (3×100 mL),respectively, and dried over sodium sulfate to provide a 3rd solution.Then the sodium sulfate was filtered and washed with ethyl acetate toprovide a 4th mixture. The 3rd solution and the 4th mixture werecombined, and evaporated to dryness. The obtained residue andBoc-L-aspartic acid (24 g) were dissolved in acetone (300 mL) to providea 5th mixture. 1-Ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride(40 g) and 4-dimethylaminopyridine (22 g) were added into the 5thmixture to provide a 6th mixture. The 6th mixture was stirred overnightat RT and evaporated to dryness. Ethyl acetate (500 ml) was added intothe residue to provide a 7th mixture, which was washed with water (2×100mL), 5% sodium bucarbonate (3×100 mL), water (100 mL), 20% citric acid(2×200 mL), and water (3×100 mL), respectively, and dried over sodiumsulfate to provide a 8th solution. The sodium sulfate was filtered andwashed with ethyl acetate to provide a 9th solution. The 8th solutionand the 9th solution were combined and evaporated to dryness. Theresidue was dissolved in methanol (R0084, 300 mL) to provide a 10thmixture. To the 10th mixture was added first palladium on activatedcharcoal (10 g, 10%) under nitrogen, and then bubbled with hydrogen gasto remove benzyloxycarbonyl group at RT. The obtained mixture (11thmixture) was filtered to remove the palladium on activated charcoal, andevaporated to dryness.

The residue was suspended in DCM (200 mL) to provide a 12th mixture.Sodium bicarbonate (20 g) and tetrabutylammonium hydrogen sulfate (11 g)were added into the 12th mixture to provide a 13th mixture. Then1-chloroethyl chloroformate (16 g) was added into the 13th mixture toprovide an 14th mixture. The 14th mixture was stirred at RT overnight.Then the organic layer of the 14th mixture was collected and washed withwater (3×200 mL), and dried over anhydrous sodium sulfate to provide a15th solution. The sodium sulfate was removed by filtration and washedwith DCM to provide a 16th solution. The 15th solution and the 16thsolution were combined and evaporated to dryness. The residue wasdissolved in isobutyric acid (100 mL) to provide a 17th mixture. Amixture of diisopropylethylamine (60 mL) and isobutyric acid (36 mL) wasprepared first and then added into the 17th mixture to provide a 18thmixture. The 18th mixture was stirred at 55° C. for 48 hours, and addedethyl acetate (500 mL) with stirring. The obtained organic layer wascollected and washed with 5% sodium bicarbonate (3×100 mL) and water(3×100 mL), respectively, and dried over anhydrous sodium sulfate toprovide a 19th solution. The sodium sulfate was filtered and washed withethyl acetate to provide a 20th solution. The 19th solution and the 20thsolution were combined and concentrated to 300 mL. Into the concentratedsolution was added anisole (20 g) and then bubbled HF gas (20 g) toprovide precipitation. The precipitated solid was collected and washedwith ethyl acetate to yield1-(((2-(4-amino-2,5-dioxo-2,3,4,5-tetrahydrobenzo[b][1,4]dioxocin-8-yl)ethyl)carbamoyl)oxy)ethylisobutyrate hydrofluoride and1-(((2-(3-amino-2,5-dioxo-2,3,4,5-tetrahydrobenzo[b][1,4]dioxocin-8-yl)ethyl)carbamoyl)oxy)ethylisobutyrate hydrofluoride.

Example 9 Preparation of4-(2-((ethoxycarbonyl)amino)ethyl)-2-(2-methylamino)acetoxy)phenylbenzoate hydrochloride and5-(2-((ethoxycarbonyl)amino)ethyl)-2-(2-methylamino)acetoxy)phenylbenzoate hydrochloride

In this example, a dopamine.HCl solution was prepared by addingdopamine.HCl (19 g) into acetone (200 mL). Pyridine (40 ml) was addedinto the dopamine.HCl solution to provide a 1st mixture. ThenN-(ethoxycarbonyloxy)succinimide (18 g) was added to the 1st mixture toprovide a 2nd mixture. The 2nd mixture was stirred at RT overnight, andadded ethyl acetate (R0061, 500 mL) for extraction. The organic layerwas separated, and washed with water (2×100 mL), 20% citric acid (2×200mL), and water (3×100 mL), respectively, and dried over sodium sulfateto provide a 3rd solution. Then the sodium sulfate was filtered andwashed with ethyl acetate to provide a 4th mixture. The 3rd solution andthe 4th mixture were combined, and evaporated to dryness. The obtainedresidue and Trt-sarcocine (36 g) were dissolved in acetone (300 mL) toprovide a 5th mixture. 1-Ethyl-(3-dimethylaminopropyl)carbodiimidehydrochloride (20 g) and 4-dimethylaminopyridine (12 g) were added intothe 5th mixture to provide a 6th mixture. The 6th mixture was stirredovernight at RT and evaporated to dryness. Ethyl acetate (500 ml) wasadded into the residue to provide a 7th mixture, which was washed withwater (2×100 mL), 5% sodium bucarbonate (3×100 mL), water (100 mL), 20%citric acid (2×200 mL), and water (3×100 mL), respectively, and driedover sodium sulfate to provide a 8th solution. The sodium sulfate wasfiltered and washed with ethyl acetate to provide a 9th solution. To thecombined 8th solution and the 9th solution were added pyridine (20 mL)and then drop by drop added benzoyl chloride (15 g) to provide a 10thmixture.

The 10th mixture was stirred at RT for 4 hours, and washed with water(2×100 mL), 5% sodium bucarbonate (2×100 mL), water (100 mL), 20% citricacid (2×200 mL), and water (3×100 mL), respectively, and dried oversodium sulfate to provide a 11th solution. The sodium sulfate wasfiltered and washed with ethyl acetate to provide a 12th solution.

Into the combined 11th and 12th solutions was added anisole (20 g) andthen bubbled HCl gas (20 g) to provide precipitation. The precipitatedsolid was collected and washed with ethyl acetate to yield4-(2-((ethoxycarbonyl)amino)ethyl)-2-(2-methylamino)acetoxy)phenylbenzoate hydrochloride and5-(2-((ethoxycarbonyl)amino)ethyl)-2-(2-methylamino)acetoxy)phenylbenzoate.

Example 10 Preparation of 4-(2-aminoethyl)-1,2-phenylene dibenzoatehydrochloride

In this example, a dopamine.HCl solution was prepared by addingdopamine.HCl (19 g) into acetone (200 mL). Water (200 mL) and sodiumbicarbonate (R0090, 50 g) were added into the dopamine.HCl solution toprovide a 1st mixture. Then di-tert-butyl dicarbonate (22 g) was addedto the 1st mixture to provide a 2nd mixture. The 2nd mixture was stirredat RT overnight, and added ethyl acetate (R0061, 500 mL) for extraction.The organic layer was separated, and washed with water (2×100 mL), 20%citric acid (2×200 mL), and water (3×100 mL), respectively, and driedover sodium sulfate to provide a 3rd solution. Then the sodium sulfatewas filtered and washed with ethyl acetate to provide a 4th mixture. The3rd solution and the 4th mixture were combined, and added first withpyridine (R0081, 30 mL), and then with benzoyl chloride (R0488, 30 g)drop by drop to provide a 5th mixture. The 5th mixture was stirred at RTfor 2 hours, then washed with water (2×100 mL), 5% sodium bucarbonate(3×100 mL), water (100 mL), 20% citric acid (2×200 mL), and water (3×100mL), respectively, and dried over sodium sulfate to provide a 6thsolution. The sodium sulfate was filtered and washed with ethyl acetateto provide a 7th solution. The 6th solution and the 7th solution werecombined and concentrated to about 300 mL (an 8th solution). Into the8th solution was added anisole (20 g), and then bubbled HCl gas (60 g)to form precipitation. The precipitated solid was collected and washedwith ethyl acetate to yield 4-(2-aminoethyl)-1,2-phenylene dibenzoatehydrochloride.

Example 11 Preparation of(S)-4-(2-amino-3-isopropoxy-3-oxopropyl)-1,2-phenylene dibenzoatehydrochloride

In this example, (S)-2-amino-3-(3,4-dihydroxyphenyl)propanoic acid (20g) was added into isopropanol (200 mL) to provide a 1st mixture. HCl gas(20 g) was bubbled into the 1st mixture to provide a 2nd mixture. The2nd mixture was stirred for 2 days at 60° C., and then added withisopropyl acetate (200 mL) to provide precipitation. The precipitatedsolid was collected and washed with isopropayl acetate to provideisopropyl (S)-2-amino-3-(3,4-dihydroxyphenyl)propanoate hydrochloride.

Isopropyl (S)-2-amino-3-(3,4-dihydroxyphenyl)propanoate hydrochloride(28 g) was added into acetone (200 mL) to provide a 3rd mixture. Intothe 3rd mixture was added water (200 mL) and sodium bicarbonate (50 g),and then di-tert-butyl dicarbonate (22 g) to provide a 4th mixture. The4th mixture was stirred for overnight at RT, and added ethyl acetate(500 mL). The organic layer (5th mixture) was washed with water (2×100mL), 20% citric acid (2×200 mL), and water (3×100 mL), respectively, anddried over anhydrous sodium sulfate to provide a 6th solution. Then thesodium sulfate was filtered and washed with ethyl acetate to provide a7th mixture. The 6th solution and the 7th mixture were combined, andadded first with pyridine (R0081, 30 mL), and then with benzoyl chloride(R0488, 30 g) drop by drop to provide an 8th mixture. The 8th mixturewas stirred at RT for 2 hours, then washed with water (2×100 mL), 5%sodium bucarbonate (3×100 mL), water (100 mL), 20% citric acid (2×200mL), and water (3×100 mL), respectively, and dried over sodium sulfateto provide a 9th solution. The sodium sulfate was filtered and washedwith ethyl acetate to provide a 10th solution. The 9th solution and the10th solution were combined and concentrated to about 300 mL (an 11thsolution). Into the 11th solution was added anisole (20 g), and thenbubbled HCl gas (20 g) to form precipitation. The precipitated solid wascollected and washed with ethyl acetate to yield(S)-4-(2-amino-3-isopropoxy-3-oxopropyl)-1,2-phenylene dibenzoatehydrochloride.

Example 12 Preparation of(S)-4-(2-amino-3-(heptan-4-yloxy)-3-oxopropyl)-1,2-phenylene dibenzoatehydrochloride

In this example, (S)-2-amino-3-(3,4-dihydroxyphenyl)propanoic acid (20g) was added into 4-heptanol (200 mL) to provide a 1st mixture. HCl gas(20 g) was bubbled into the 1st mixture to provide a 2nd mixture. The2nd mixture was stirred for 2 days at 60° C., and then added withisopropyl acetate (200 mL) to provide precipitation. The precipitatedsolid was collected and washed with isopropayl acetate to provide4-heptyl (S)-2-amino-3-(3,4-dihydroxyphenyl)propanoate hydrochloride.

4-Heptyl (S)-2-amino-3-(3,4-dihydroxyphenyl)propanoate hydrochloride (28g) was added into acetone (200 mL) to provide a 3rd mixture. Into the3rd mixture was added water (200 mL) and sodium bicarbonate (50 g), andthen di-tert-butyl dicarbonate (22 g) to provide a 4th mixture. The 4thmixture was stirred for overnight at RT, and added ethyl acetate (500mL). The organic layer (5th mixture) was washed with water (2×100 mL),20% citric acid (2×200 mL), and water (3×100 mL), respectively, anddried over anhydrous sodium sulfate to provide a 6th solution. Then thesodium sulfate was filtered and washed with ethyl acetate to provide a7th mixture. The 6th solution and the 7th mixture were combined, andadded first with pyridine (R0081, 30 mL), and then with benzoyl chloride(R0488, 30 g) drop by drop to provide an 8th mixture. The 8th mixturewas stirred at RT for 2 hours, then washed with water (2×100 mL), 5%sodium bucarbonate (3×100 mL), water (100 mL), 20% citric acid (2×200mL), and water (3×100 mL), respectively, and dried over sodium sulfateto provide a 9th solution. The sodium sulfate was filtered and washedwith ethyl acetate to provide a 10th solution. The 9th solution and the10th solution were combined and concentrated to about 300 mL (an 11thsolution). Into the 11th solution was added anisole (20 g), and thenbubbled HCl gas (20 g) to form precipitation. The precipitated solid wascollected and washed with ethyl acetate to yield(S)-4-(2-amino-3-(heptan-4-yloxy)-3-oxopropyl)-1,2-phenylene dibenzoatehydrochloride.

Example 13 Preparation of(S)-4-(2-amino-3-isopropoxy-3-oxopropyl)-1,2-phenylene dipentanoatehydrochloride

In this example, (S)-2-amino-3-(3,4-dihydroxyphenyl)propanoic acid (20g) was added into isopropanol (200 mL) to provide a 1st mixture. HCl gas(20 g) was bubbled into the 1st mixture to provide a 2nd mixture. The2nd mixture was stirred for 2 days at 60° C., and then added withisopropyl acetate (200 mL) to provide precipitation. The precipitatedsolid was collected and washed with isopropayl acetate to provideisopropyl (S)-2-amino-3-(3,4-dihydroxyphenyl)propanoate hydrochloride.

Isopropyl (S)-2-amino-3-(3,4-dihydroxyphenyl)propanoate hydrochloride(28 g) was added into acetone (200 mL) to provide a 3rd mixture. Intothe 3rd mixture was added water (200 mL) and sodium bicarbonate (50 g),and then di-tert-butyl dicarbonate (22 g) to provide a 4th mixture. The4th mixture was stirred for overnight at RT, and added ethyl acetate(500 mL). The organic layer (5th mixture) was washed with water (2×100mL), 20% citric acid (2×200 mL), and water (3×100 mL), respectively, anddried over anhydrous sodium sulfate to provide a 6th solution. Then thesodium sulfate was filtered and washed with ethyl acetate to provide a7th mixture. The 6th solution and the 7th mixture were combined, andadded first with pyridine (R0081, 30 mL), and then with pentanoylchloride (24 g) drop by drop to provide an 8th mixture. The 8th mixturewas stirred at RT for 2 hours, then washed with water (2×100 mL), 5%sodium bucarbonate (3×100 mL), water (100 mL), 20% citric acid (2×200mL), and water (3×100 mL), respectively, and dried over sodium sulfateto provide a 9th solution. The sodium sulfate was filtered and washedwith ethyl acetate to provide a 10th solution. The 9th solution and the10th solution were combined and concentrated to about 300 mL (an 11thsolution). Into the 11th solution was added anisole (20 g), and thenbubbled HCl gas (20 g) to form precipitation. The precipitated solid wascollected and washed with ethyl acetate to yield(S)-4-(2-amino-3-isopropoxy-3-oxopropyl)-1,2-phenylene dipentanoatehydrochloride.

Example 14 Preparation of(S)-4-(2-amino-3-ethoxy-3-oxopropyl)-1,2-phenylene diacetatehydrochloride

In this example, (S)-2-amino-3-(3,4-dihydroxyphenyl)propanoic acid (20g) was added into ethanol (200 mL) to provide a 1st mixture. HCl gas (20g) was bubbled into the 1st mixture to provide a 2nd mixture. The 2ndmixture was stirred for 2 days at 60° C., and then added with isopropylacetate (200 mL) to provide precipitation. The precipitated solid wascollected and washed with isopropayl acetate to provide ethyl(S)-2-amino-3-(3,4-dihydroxyphenyl)propanoate hydrochloride.

Ethyl (S)-2-amino-3-(3,4-dihydroxyphenyl)propanoate hydrochloride (26 g)was added into acetone (200 mL) to provide a 3rd mixture. Into the 3rdmixture was added water (200 mL) and sodium bicarbonate (50 g), and thendi-tert-butyl dicarbonate (22 g) to provide a 4th mixture. The 4thmixture was stirred for overnight at RT, and added ethyl acetate (500mL). The organic layer (5th mixture) was washed with water (2×100 mL),20% citric acid (2×200 mL), and water (3×100 mL), respectively, anddried over anhydrous sodium sulfate to provide a 6th solution. Then thesodium sulfate was filtered and washed with ethyl acetate to provide a7th mixture. The 6th solution and the 7th mixture were combined, andadded first with pyridine (R0081, 30 mL), and then with acetyl chloride(18 g) drop by drop to provide an 8th mixture. The 8th mixture wasstirred at RT for 2 hours, then washed with water (2×100 mL), 5% sodiumbucarbonate (3×100 mL), water (100 mL), 20% citric acid (2×200 mL), andwater (3×100 mL), respectively, and dried over sodium sulfate to providea 9th solution. The sodium sulfate was filtered and washed with ethylacetate to provide a 10th solution. The 9th solution and the 10thsolution were combined and concentrated to about 300 mL (an 11thsolution). Into the 11th solution was added anisole (20 g), and thenbubbled HCl gas (20 g) to form precipitation. The precipitated solid wascollected and washed with ethyl acetate to yield(S)-4-(2-amino-3-ethoxy-3-oxopropyl)-1,2-phenylene diacetatehydrochloride.

Example 15 Preparation of(S)-4-(2-amino-3-oxo-3-(pentan-3-yloxy)propyl)-1,2-phenylenebis(2-methylpropanoate) hydrobromide

In this example, (S)-2-amino-3-(3,4-dihydroxyphenyl)propanoic acid (20g) was added into 3-pentanol (200 mL) to provide a 1st mixture. HCl gas(20 g) was bubbled into the 1st mixture to provide a 2nd mixture. The2nd mixture was stirred for 2 days at 60° C., and then added withisopropyl acetate (200 mL) to provide precipitation. The precipitatedsolid was collected and washed with isopropayl acetate to provideisopropyl (S)-2-amino-3-(3,4-dihydroxyphenyl)propanoate hydrochloride.

3-Pentyl (S)-2-amino-3-(3,4-dihydroxyphenyl)propanoate hydrochloride (28g) was added into acetone (200 mL) to provide a 3rd mixture. Into the3rd mixture was added water (200 mL) and sodium bicarbonate (50 g), andthen di-tert-butyl dicarbonate (22 g) to provide a 4th mixture. The 4thmixture was stirred for overnight at RT, and added ethyl acetate (500mL). The organic layer (5th mixture) was washed with water (2×100 mL),20% citric acid (2×200 mL), and water (3×100 mL), respectively, anddried over anhydrous sodium sulfate to provide a 6th solution. Then thesodium sulfate was filtered and washed with ethyl acetate to provide a7th mixture. The 6th solution and the 7th mixture were combined, andadded first with pyridine (R0081, 30 mL), and then with isobutyrylchloride (22 g) drop by drop to provide an 8th mixture. The 8th mixturewas stirred at RT for 2 hours, then washed with water (2×100 mL), 5%sodium bucarbonate (3×100 mL), water (100 mL), 20% citric acid (2×200mL), and water (3×100 mL), respectively, and dried over sodium sulfateto provide a 9th solution. The sodium sulfate was filtered and washedwith ethyl acetate to provide a 10th solution. The 9th solution and the10th solution were combined and concentrated to about 300 mL (an 11thsolution). Into the 11th solution was added anisole (20 g), and thenbubbled HCl gas (20 g) to form precipitation. The precipitated solid wascollected and washed with ethyl acetate to yield(S)-4-(2-amino-3-ethoxy-3-oxopropyl)-1,2-phenylene diacetatehydrochloride.

Example 16 Preparation of(S)-4-(2-aminoacetamido)-3-isopropoxy-3-oxopropyl)-1,2-phenylenedibenzoate hydrochloride

In this example, (S)-2-amino-3-(3,4-dihydroxyphenyl)propanoic acid (20g) was added into isopropanol (200 mL) to provide a 1st mixture. HCl gas(20 g) was bubbled into the 1st mixture to provide a 2nd mixture. The2nd mixture was stirred for 2 days at 60° C., and then added withisopropyl acetate (200 mL) to provide precipitation. The precipitatedsolid was collected and washed with isopropayl acetate to provideisopropyl (S)-2-amino-3-(3,4-dihydroxyphenyl)propanoate hydrochloride.

Isopropyl (S)-2-amino-3-(3,4-dihydroxyphenyl)propanoate hydrochloride(28 g) was added into acetone (200 mL) to provide a 3rd mixture. Intothe 3rd mixture was added water (200 mL) and sodium bicarbonate (50 g),and then N-(tert-Butoxycarbonyl-glycine N-hydroxysuccinimide ester (27g) to provide a 4th mixture. The 4th mixture was stirred for overnightat RT, and added ethyl acetate (500 mL). The organic layer (5th mixture)was washed with water (2×100 mL), 20% citric acid (2×200 mL), and water(3×100 mL), respectively, and dried over anhydrous sodium sulfate toprovide a 6th solution. Then the sodium sulfate was filtered and washedwith ethyl acetate to provide a 7th mixture. The 6th solution and the7th mixture were combined, and added first with pyridine (R0081, 30 mL),and then with benzoyl chloride (30 g) drop by drop to provide an 8thmixture. The 8th mixture was stirred at RT for 2 hours, then washed withwater (2×100 mL), 5% sodium bucarbonate (3×100 mL), water (100 mL), 20%citric acid (2×200 mL), and water (3×100 mL), respectively, and driedover sodium sulfate to provide a 9th solution. The sodium sulfate wasfiltered and washed with ethyl acetate to provide a 10th solution. The9th solution and the 10th solution were combined and concentrated toabout 300 mL (an 11th solution). Into the 11th solution was addedanisole (20 g), and then bubbled HCl gas (20 g) to form precipitation.The precipitated solid was collected and washed with ethyl acetate toyield (S)-4-(2-aminoacetamido)-3-isopropoxy-3-oxopropyl)-1,2-phenylenedibenzoate hydrochloride.

Example 17 Preparation of4-((2S)-3-oxo-3-(pentan-3-yloxy)-2-(pyrrolidine-2-carboxamido)propyl)-1,2-phenylenedibenzoate hydrofluoride

In this example, (S)-2-amino-3-(3,4-dihydroxyphenyl)propanoic acid (20g) was added into 3-pentanol (200 mL) to provide a 1st mixture. HCl gas(20 g) was bubbled into the 1st mixture to provide a 2nd mixture. The2nd mixture was stirred for 2 days at 60° C., and then added withisopropyl acetate (200 mL) to provide precipitation. The precipitatedsolid was collected and washed with isopropayl acetate to provideisopropyl (S)-2-amino-3-(3,4-dihydroxyphenyl)propanoate hydrochloride.

3-Pentyl (S)-2-amino-3-(3,4-dihydroxyphenyl)propanoate hydrochloride (28g) was added into acetone (200 mL) to provide a 3rd mixture. Into the3rd mixture was added water (200 mL) and sodium bicarbonate (50 g), andthen N-(tert-Butoxycarbonyl-proline N-hydroxysuccinimide ester (32 g) toprovide a 4th mixture. The 4th mixture was stirred for overnight at RT,and added ethyl acetate (500 mL). The organic layer (5th mixture) waswashed with water (2×100 mL), 20% citric acid (2×200 mL), and water(3×100 mL), respectively, and dried over anhydrous sodium sulfate toprovide a 6th solution. Then the sodium sulfate was filtered and washedwith ethyl acetate to provide a 7th mixture. The 6th solution and the7th mixture were combined, and added first with pyridine (R0081, 30 mL),and then with benzoyl chloride (22 g) drop by drop to provide an 8thmixture. The 8th mixture was stirred at RT for 2 hours, then washed withwater (2×100 mL), 5% sodium bucarbonate (3×100 mL), water (100 mL), 20%citric acid (2×200 mL), and water (3×100 mL), respectively, and driedover sodium sulfate to provide a 9th solution. The sodium sulfate wasfiltered and washed with ethyl acetate to provide a 10th solution. The9th solution and the 10th solution were combined and concentrated toabout 300 mL (an 11th solution). Into the 11th solution was addedanisole (20 g), and then bubbled HF gas (20 g) to form precipitation.The precipitated solid was collected and washed with ethyl acetate toyield4-((2S)-3-oxo-3-(pentan-3-yloxy)-2-(pyrrolidine-2-carboxamido)propyl)-1,2-phenylenedibenzoate hydrofluoride.

Example 18 Preparation of(S)-4-(3-isopropoxy-3-oxo-2-(piperidine-4-carboxamido)propyl)-1,2-phenylenedibenzoate hydrofluoride

In this example, (S)-2-amino-3-(3,4-dihydroxyphenyl)propanoic acid (20g) was added into isopropanol (200 mL) to provide a 1st mixture. HCl gas(20 g) was bubbled into the 1st mixture to provide a 2nd mixture. The2nd mixture was stirred for 2 days at 60° C., and then added withisopropyl acetate (200 mL) to provide precipitation. The precipitatedsolid was collected and washed with isopropayl acetate to provideisopropyl (S)-2-amino-3-(3,4-dihydroxyphenyl)propanoate hydrochloride.

Isopropyl (S)-2-amino-3-(3,4-dihydroxyphenyl)propanoate hydrochloride(28 g) was added into acetone (200 mL) to provide a 3rd mixture. Intothe 3rd mixture was added water (200 mL) and sodium bicarbonate (50 g),and then Boc-piperidine-4-carboxylic acid N-hydroxysuccinimide ester(Boc-Inp-OSu, 33 g) to provide a 4th mixture. The 4th mixture wasstirred for overnight at RT, and added ethyl acetate (500 mL). Theorganic layer (5th mixture) was washed with water (2×100 mL), 20% citricacid (2×200 mL), and water (3×100 mL), respectively, and dried overanhydrous sodium sulfate to provide a 6th solution. Then the sodiumsulfate was filtered and washed with ethyl acetate to provide a 7thmixture. The 6th solution and the 7th mixture were combined, and addedfirst with pyridine (R0081, 30 mL), and then with benzoyl chloride (30g) drop by drop to provide an 8th mixture. The 8th mixture was stirredat RT for 2 hours, then washed with water (2×100 mL), 5% sodiumbucarbonate (3×100 mL), water (100 mL), 20% citric acid (2×200 mL), andwater (3×100 mL), respectively, and dried over sodium sulfate to providea 9th solution. The sodium sulfate was filtered and washed with ethylacetate to provide a 10th solution. The 9th solution and the 10thsolution were combined and concentrated to about 300 mL (an 11thsolution). Into the 11th solution was added anisole (20 g), and thenbubbled HF gas (20 g) to form precipitation. The precipitated solid wascollected and washed with ethyl acetate to yield(S)-4-(3-isopropoxy-3-oxo-2-(piperidine-4-carboxamido)propyl)-1,2-phenylenedibenzoate hydrofluoride.

Example 19 Preparation of4-((2S)-3-isopropoxy-3-2-(octahydro-1H-quinolizine-2-carboxamido)-3-oxopropyl)-1,2-phenylenebis(2-methylpropanoate) hydrochloride

In this example, (S)-2-amino-3-(3,4-dihydroxyphenyl)propanoic acid (20g) was added into isopropanol (200 mL) to provide a 1st mixture. HCl gas(20 g) was bubbled into the 1st mixture to provide a 2nd mixture. The2nd mixture was stirred for 2 days at 60° C., and then added withisopropyl acetate (200 mL) to provide precipitation. The precipitatedsolid was collected and washed with isopropayl acetate to provideisopropyl (S)-2-amino-3-(3,4-dihydroxyphenyl)propanoate hydrochloride.

Isopropyl (S)-2-amino-3-(3,4-dihydroxyphenyl)propanoate hydrochloride(28 g) was added into acetone (200 mL) to provide a 3rd mixture. Intothe 3rd mixture was added water (200 mL) and sodium bicarbonate (50 g),and then octahydro-1H-quinolizine-2-carboxylic acid N-hydroxysuccinimideester (28 g) to provide a 4th mixture. The 4th mixture was stirred forovernight at RT, and added ethyl acetate (500 mL). The organic layer(5th mixture) was washed with water (2×100 mL), 20% citric acid (2×200mL), and water (3×100 mL), respectively, and dried over anhydrous sodiumsulfate to provide a 6th solution. Then the sodium sulfate was filteredand washed with ethyl acetate to provide a 7th mixture. The 6th solutionand the 7th mixture were combined, and added first with pyridine (R0081,30 mL), and then with isobutyryl chloride (22 g) drop by drop to providean 8th mixture. The 8th mixture was stirred at RT for 2 hours, thenwashed with water (2×100 mL), 5% sodium bucarbonate (3×100 mL), water(100 mL), 20% citric acid (2×200 mL), and water (3×100 mL),respectively, and dried over sodium sulfate to provide a 9th solution.The sodium sulfate was filtered and washed with ethyl acetate to providea 10th solution. The 9th solution and the 10th solution were combinedand concentrated to about 100 mL (an 11th solution). Into the 11thsolution was added hexane (200 mL), and then acetic acid (6 g) to formprecipitation. The precipitated solid was collected and washed withethyl acetate/hexane to4-((2S)-3-isopropoxy-3-2-(octahydro-1H-quinolizine-2-carboxamido)-3-oxopropyl)-1,2-phenylenebis(2-methylpropanoate) hydrochloride.

Example 20 Preparation of (2S)-isopropyl3-(3-amino-2,5-dioxo-2,3,4,5-tetrahydrobenzo[b][1,4]dioxocin-8-yl)-2-(((1-(isobutyryloxy)ethoxy)carbonyl)amino)propanoatehydrobromide and (2S)-isopropyl3-(4-amino-2,5-dioxo-2,3,4,5-tetrahydrobenzo[b][1,4]dioxocin-8-yl)-2-(((1-(isobutyryloxy)ethoxy)carbonyl)amino)propanoatehydrobromide

In this example, (S)-2-amino-3-(3,4-dihydroxyphenyl)propanoic acid (20g) was added into isopropanol (200 mL) to provide a 1st mixture. HCl gas(20 g) was bubbled into the 1st mixture to provide a 2nd mixture. The2nd mixture was stirred for 2 days at 60° C., and then added withisopropyl acetate (200 mL) to provide precipitation. The precipitatedsolid was collected and washed with isopropayl acetate to provideisopropyl (S)-2-amino-3-(3,4-dihydroxyphenyl)propanoate hydrochloride.

An isopropyl (S)-2-amino-3-(3,4-dihydroxyphenyl)propanoate hydrochloridesolution was prepared by adding isopropyl(S)-2-amino-3-(3,4-dihydroxyphenyl)propanoate hydrochloride (28 g) intoacetone (200 mL). Water (200 mL) and sodium bicarbonate (R0090, 50 g)were added into the dopamine.HCl solution to provide a 3rd mixture. ThenNα-(benzyloxycarbonyloxy)succinimide (25 g) was added to the 3rd mixtureto provide a 4th mixture. The 4th mixture was stirred at RT overnight,and added ethyl acetate (R0061, 500 mL) for extraction. The organiclayer was separated, and washed with water (2×100 mL), 20% citric acid(2×200 mL), and water (3×100 mL), respectively, and dried over sodiumsulfate to provide a 5th solution. Then the sodium sulfate was filteredand washed with ethyl acetate to provide a 6th solution. The 5thsolution and the 6th mixture were combined, and evaporated to dryness.The obtained residue and Boc-L-aspartic acid (24 g) were dissolved inacetone (300 mL) to provide a 7th mixture.1-Ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (40 g) and4-dimethylaminopyridine (22 g) were added into the 7th mixture toprovide an 8th mixture. The 8th mixture was stirred overnight at RT andevaporated to dryness. Ethyl acetate (500 ml) was added into the residueto provide a 9th mixture, which was washed with water (2×100 mL), 5%sodium bucarbonate (3×100 mL), water (100 mL), 20% citric acid (2×200mL), and water (3×100 mL), respectively, and dried over sodium sulfateto provide a 10th solution. The sodium sulfate was filtered and washedwith ethyl acetate to provide an 11th solution. The 10th solution andthe 11th solution were combined and evaporated to dryness. The residuewas dissolved in methanol (R0084, 300 mL) to provide a 12th mixture. Tothe 12th mixture was added first palladium on activated charcoal (10 g,10%) under nitrogen, and then bubbled with hydrogen gas to removebenzyloxycarbonyl group at RT. The obtained mixture (13th mixture) wasfiltered to remove the palladium on activated charcoal, and evaporatedto dryness.

The residue was suspended in DCM (200 mL) to provide a 14th mixture.Sodium bicarbonate (15 g) and tetrabutylammonium hydrogen sulfate (11 g)were added into the 14th mixture to provide a 15th mixture. Then1-chloroethyl chloroformate (16 g) was added into the 15th mixture toprovide a 16th mixture. The 16th mixture was stirred at RT overnight.Then the organic layer of the 16th mixture was collected and washed withwater (3×200 mL), and dried over anhydrous sodium sulfate to provide a17th solution. The sodium sulfate was removed by filtration and washedwith DCM to provide a 18th solution. The 17th solution and the 18thsolution were combined and evaporated to dryness. The residue wasdissolved in isobutyric acid (100 mL) to provide a 19th mixture. Amixture of diisopropylethylamine (60 mL) and isobutyric acid (36 mL) wasprepared first and then added into the 19th mixture to provide a 20thmixture. The 20th mixture was stirred at 55° C. for 48 hours, and addedethyl acetate (500 mL) with stirring. The obtained organic layer wascollected and washed with 5% sodium bicarbonate (3×100 mL) and water(3×100 mL), respectively, and dried over anhydrous sodium sulfate toprovide a 21st solution. The sodium sulfate was filtered and washed withethyl acetate to provide a 22nd solution. The 21st solution and the 22ndsolution were combined and concentrated to 300 mL. Into the concentratedsolution was added anisole (20 g) and then bubbled HBr gas (30 g) toprovide precipitation. The precipitated solid was collected and washedwith ethyl acetate to yield (2S)-isopropyl3-(3-amino-2,5-dioxo-2,3,4,5-tetrahydrobenzo[b][1,4]dioxocin-8-yl)-2-(((1-(isobutyryloxy)ethoxy)carbonyl)amino)propanoatehydrobromide and (2S)-isopropyl3-(4-amino-2,5-dioxo-2,3,4,5-tetrahydrobenzo[b][1,4]dioxocin-8-yl)-2-(((1-(isobutyryloxy)ethoxy)carbonyl)amino)propanoatehydrobromide.

Example 21 Preparation of5-((2S)-2-(((1-(isobutyryloxy)ethoxy)carbonyl)amino-3-isopropoxy-3-oxopropyl)-2-(2-(methylamino)acetoxy)phenylbenzoate hydrochloride and4-((2S)-2-(((1-(isobutyryloxy)ethoxy)carbonyl)amino-3-isopropoxy-3-oxopropyl)-2-(2-(methylamino)acetoxy)phenylbenzoate hydrochloride

In this example, (S)-2-amino-3-(3,4-dihydroxyphenyl)propanoic acid (20g) was added into isopropanol (200 mL) to provide a 1st mixture. HCl gas(20 g) was bubbled into the 1st mixture to provide a 2nd mixture. The2nd mixture was stirred for 2 days at 60° C. and then added withisopropyl acetate (200 mL) to provide precipitation. The precipitatedsolid was collected and washed with isopropayl acetate to provideisopropyl (S)-2-amino-3-(3,4-dihydroxyphenyl)propanoate hydrochloride.

An isopropyl (S)-2-amino-3-(3,4-dihydroxyphenyl)propanoate hydrochloridesolution was prepared by adding isopropyl(S)-2-amino-3-(3,4-dihydroxyphenyl)propanoate hydrochloride (28 g) intoacetone (200 mL). Water (200 mLl) and sodium bicarbonate (R0090, 50 g)were added into the dopamine.HCl solution to provide a 3rd mixture. ThenNα-(benzyloxycarbonyloxy)succinimide (25 g) was added to the 3rd mixtureto provide a 4th mixture. The 4th mixture was stirred at RT overnight,and added ethyl acetate (R0061, 500 mL) for extraction. The organiclayer was separated, and washed with water (2×100 mL), 20% citric acid(2×200 mL), and water (3×100 mL), respectively, and dried over sodiumsulfate to provide a 5th solution. Then the sodium sulfate was filteredand washed with ethyl acetate to provide a 6th solution. The 5thsolution and the 6th mixture were combined, and evaporated to dryness.

The obtained residue and Trt-sarcocine (36 g) were dissolved in acetone(300 mL) to provide a 7th mixture.1-Ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (20 g) and4-dimethylaminopyridine (12 g) were added into the 7th mixture toprovide an 8th mixture. The 8th mixture was stirred overnight at RT andevaporated to dryness. Ethyl acetate (500 ml) was added into the residueto provide a 9th mixture, which was washed with water (2×100 mL), 5%sodium bucarbonate (3×100 mL), water (100 mL), 20% citric acid (2×200mL), and water (3×100 mL), respectively, and dried over sodium sulfateto provide a 10th solution. The sodium sulfate was filtered and washedwith ethyl acetate to provide an 11th solution. Pyridine (20 ml) wasadded into the 11th solution to provide a 12th mixture, and benzoylchloride (15 g) was added into the 12th mixture drop by drop to providea 13th. mixture. The 13th. mixture was stirred for 4 hours at RT and theobtained mixture was washed with water (2×100 ml), 5% sodium bucarbonate(2×100 ml), water (100 ml), 20% citric acid (2×200 ml), and water (3×100ml), respectively, and dried over anhydrous sodium sulfate to provide a14th solution. The sodium sulfate was removed by filtration and washedwith ethyl acetate to provide a 15th solution. The 14th and the 15thsolutions were combined and evaporated to dryness.

The obtained residue was dissolved in methanol (300 mL) to provide a16th mixture. 10% Palladium on activated charcoal (10 g) was added intothe 16th mixture under nitrogen, and hydrogen gas was bubbled into theobtained mixture until the benzyloxycarbonyl group was removedsubstantially completely at RT. The palladium on activated charcoal wasremoved by filtration, and the obtained solution was evaporated todryness.

The obtained residue was suspended in DCM (200 mL). Sodium bicarbonate(15 g) and tetrabutylammonium hydrogen sulfate (11 g) were added intothe DCM suspension. 1-chloroethyl chloroformate (16 g) was added intothe obtained reaction mixture. Then the reaction mixture was stirred forovernight at RT. The organic layer was collected and washed with water(3×200 mL). The solution was dried over anhydrous sodium sulfate. Sodiumsulfate is removed by filtration and washed with DCM. The DCM solutionis evaporated to dryness.

The residue was dissolved in isobutyric acid (100 mL). A mixture of[diisopropylethylamine (60 ml) and isobutyric acid (R0874, 36 mL) whichis mixed before addition] is added into the reaction solution. Themixture is stirred for 48 h at 55 C. Ethyl acetate (500 ml) is addedinto the reaction mixture with stirring. The organic solution iscollected and washed with 5% sodium bicarbonate (3×100 ml) and water(3×100 ml). The solution is dried over anhydrous sodium sulfate. Sodiumsulfate is removed by filtration and washed with ethyl acetate (3×). Thesolution is concentrated to 300 ml. Anisole (20 g) is added into theethyl acetate solution. Then HCl gas (30 g) is bubbled into the ethylacetate solution. The solid is collected and washed with ethyl acetate.

Example 22 Preparation of4-((2S)-3-isopropoxy-2-((((octahydroindolizin-1-yl)oxy)carbonyl)amino)-3-oxopropyl)-1,2-phenylenedibenzoate acetate

In this example, (S)-2-amino-3-(3,4-dihydroxyphenyl)propanoic acid (20g) was added into isopropanol (200 mL). HCl gas (20 g) was bubbled intothe mixture. The mixture was stirred for 2 days at 60° C. Isopropylacetate (200 mL) was added into the mixture. The solid was collected andwashed with isopropayl acetate.

Isopropyl (S)-2-amino-3-(3,4-dihydroxyphenyl)propanoate hydrochloride(28 g) was added into acetone (200 mL). Water (200 mL) and sodiumbicarbonate (50 g) were added into the reaction mixture. Di-tert-butyldicarbonate (22 g) was added into the reaction mixture. The mixture wasstirred for overnight at RT. Ethyl acetate (500 mL) was added into themixture. The mixture was washed with water (2×100 mL), 20% citric acid(2×200 mL), and water (3×100 mL). The solution was dried over sodiumsulfate. Sodium sulfate was removed by filtration and washed with ethylacetate. Pyridine (30 mL) was added into the ethyl acetate solution.Benzoyl chloride (30 g) was added into the reaction mixture drop bydrop. The solution was stirred for 2 hours at RT. The solution waswashed with water (2×100 mL), 5% sodium bicarbonate (2×100 mL), water(100 mL), 20% citric acid (2×200 mL), and water (3×100 mL). The solutionwas dried over anhydrous sodium sulfate. Sodium sulfate was removed byfiltration and washed with ethyl acetate. The solution was concentratedto 200 mL. Anisole (20 g) was added into the ethyl acetate solution.Then HCl gas (20 g) was bubbled into the ethyl acetate solution. Thesolid was collected and washed with ethyl acetate.

The solid was suspended in ethyl acetate (200 mL), triethylamine (25 mL)was added into the mixture. Octahydroindolizin-1-yl carbonochloridatehydrochloride (22 g) in ethyl acetate (50 mL) was added into thereaction mixture drop by drop. The mixture was stirred for 2 hours atRT. The mixture was washed with 5% sodium bicarbonate and water (3×).Acetic acid (6 g) was added into the mixture. Hexanes (200 mL) was addedinto the mixture. The solid was collected by filtration and washed withethyl acetate/hexanes.

Example 23 Preparation of 2-(diethylamino)ethyl2-[(2,6-dichloro-3-methylphenyl)amino]benzoate.acetate

29.6 g (0.1 mol) of 2-[(2,6-dichloro-3-methylphenyl)amino]benzoic acidwas dissolved in 300 mL of chloroform. 20.6 g ofN,N′-Dicyclohexylcarbodiimide was added into the reaction mixture. 11.7g of diethylaminoethylamine was added into the reaction mixture. Themixture was stirred for 3 hours at RT. The solid was removed byfiltration. The chloroform solution was washed with 5% NaHCO₃ (2×100 mL)and water (3×100 mL). The organic solution was dried over anhydroussodium sulfate. Sodium sulfate was removed by filtration. 6 g of aceticacid was added into the reaction mixture with stirring. Hexane (200 ml)was added. The solid product was collected by filtration.

Example 24 Preparation of (Z)-2-(diethylaminoethyl)ethyl2-(5-fluoro-2-methyl-1-(4-methylsulfinyl)benzylidene)-1H-inden-1-yl)acetate.AcOH

11.7 g (0.1 mol) of diethylaminoethanol was dissolved in 10% sodiumbicarbonate (200 mL) and acetone (100 mL). 37.5 g (0.1 mol) of(Z)-5-fluoro-2-methyl-1-[(4-methylsulfinyl)phenylmethylene]-1H-indene-3-acetylchloride was added into the reaction mixture. The mixture was stirredfor 3 hours at RT. The solvents were evaporated off. The residue wassuspended in ethyl acetate (500 mL). 5% sodium bicarbonate (200 mL) wasadded into the reaction mixture with stirring. Ethyl acetate layer wascollected and washed with water (3×500 mL). The ethyl acetate solutionwas dried over anhydrous sodium sulfate. Sodium sulfate was removed byfiltration. 6 g of acetic acid was added into the reaction mixture withstirring. The organic solution was evaporated off.

Example 25 Preparation of 2-(dimethylamino)ethyl 2-(3-phenoxyphenyl)propionate.hydrochloride

26.1 g (0.1 mol) of 2-(3-phenoxyphenyl) propionyl chloride was dissolvedin 300 ml of ethyl acetate. The mixture was cooled to 0° C. 8.9 g ofdimethylaminoethanol were added into the reaction mixture. Sodiumbicarbonate (30 g) was added into the mixture. The mixture was stirredfor 5 hours at RT. The mixture was washed with water (3×200 mL). Theethyl acetate solution was dried over anhydrous sodium sulfate. HCl gas(5 g) was bubbled into the mixture. The solid was collected byfiltration and washed with ethyl acetate.

Example 26 Preparation of S-(2-(dimethylamino)ethyl 2-(3-phenoxyphenyl)propanethioate hydrochloride

10.4 g (0.1 mol) of dimethylaminoethyl mercaptan was dissolved in 10%sodium bicarbonate (200 mL) and acetone (100 mL). 27.3 g (0.1 mol) of2-(3-phenoxyphenyl) propionyl chloride was added into the reactionmixture. The mixture was stirred for 3 hours at RT. The solvents wereevaporated off. The residue was suspended in ethyl acetate (500 mL). 5%sodium bicarbonate (200 mL) was added into the reaction mixture withstirring. Ethyl acetate layer was collected and washed with water (3×500mL). The ethyl acetate solution was dried over anhydrous sodium sulfate.Sodium sulfate was removed by filtration. Anhydrous HCl gas (5 g) wasbubbled into the reaction mixture with stirring. The solid was collectedand washed with ethyl acetate.

Example 27 Preparation of 2-(dipropylamino)ethyl4-acetoxy-2′,4′-difluoro-[1,1′-biphenyl]-3-carboxylatehydrochloride[2-(dipropylamino)ethyl5-(2,4-difluorophenyl)acetylsalicylate hydrochloride]

31.1 g (0.1 mol) of 5-(2,4-difluorophenyl) acetylsalicyl chloride wasdissolved in 300 mL of ethyl acetate. The mixture was cooled to 0° C.11.7 g (0.1 mol) of diethylaminoethanol were added into the reactionmixture. The sodium bicarbonate (30 g) was added into the reactionmixture. The mixture was stirred for 3 hours at RT. Then water (200 mL)was added into the mixture. The ethyl acetate layer was collected andwashed with water (3×). The solution was dried over anhydrous sodiumsulfate. Anhydrous HCl gas was bubbled into the reaction mixture withstirring. The solid was collected and washed with ethyl acetate.

Example 28 Preparation of 2-(diethylamino)ethyl2-(4-isobutylphenyl)propionate hydrochloride

41 g of ibuprofen was dissolved in 200 ml of ethyl acetate. 16 mL ofthionyl chloride was added into the mixture. The mixture was refluxedfor 2 h. The mixture was evaporated to dryness completely. 500 mL ofethyl acetate was added into the residue and evaporated off. 500 mL ofethyl acetate was added into the reaction mixture. The solution wascooled to 5° C. with ice-water bath. 23 g of N,N-diethylaminoethanol wasadded into the reaction mixture drop by drop. 40 g of Na₂CO₃ was addedinto the reaction mixture slowly. The mixture was stirred for overnightat RT. 200 mL of water was added into the mixture. The ethyl acetatesolution was collected and washed with water (3×200 mL) and dried overanhydrous Na₂SO₄. Sodium sulfate was removed by filtration and washedwith ethyl acetate (3×100 mL). Anhydrous HCl gas (10 g) was bubbled intothe mixture. The solid was collected and washed with ethyl acetate.

Example 29 Preparation of 2-(diethylamino)ethyl 2-acetoxybenzoatehydrochloride (2-(diethylamino)ethyl acetylsalicylate hydrochloride)

36 g of aspirin was dissolved in 100 mL of ethyl acetate. 16 mL ofthionyl chloride was added into the mixture. The mixture was refluxedfor 3 h. The mixture was evaporated to dryness completely. 100 mL ofethyl acetate was added into the residue and evaporated off. 500 mL ofethyl acetate was added into the reaction mixture. The solution wascooled to 5° C. with ice-water bath. 23 g of N-diethylaminoethanol wasadded into the reaction mixture drop by drop. 40 g of NaHCO₃ was addedinto the reaction mixture slowly. The mixture was stirred for overnightat RT. 200 mL of water was added into the mixture. The ethyl acetatesolution was collected and washed with water (3×100 mL) and dried overanhydrous Na₂SO₄. Sodium sulfate was removed by filtration and washedwith ethyl acetate (3×100 mL). Anhydrous HCl gas (10 g) was bubbled intothe mixture. The solid was collected and washed with ethyl acetate.

Example 30 Measurements of Penetration Rates of HPPs

The penetrate rates of4-(2-(2-(methylamino)acetamido)ethyl)-1,2,-phenylene dibenzoatehydrochloride (Compound-1),4-(2-(2-(methylamino)acetamido)ethyl)-1,2,-phenylene dibenzoate aceticacid (Compound-2),4-(6-methyl-4,8-dioxo-5,7-dioxa-2,9-diazadecan-11-yl)-1,2,-phenylenedibenzoate hydrobromide (Compound-3),4-(2-(2-amino-3-phenylpropanamido)ethyl)-1,2,-phenylene dibenzoatehydrochloride (Compound-4),4-(2-(((1-((pyrrolidine-2-carbonyl)oxy)ethoxy)amino)ethyl)-1,2,-phenylenedibenzoate hydrochloride (Compound-5),4-(2-piperidine-4-carboxamido)ethyl)-1,2,-phenylenebis(2-ethylbutanoate) hydrochloride (Compound-6),4-(2-((((-octahydro-1H-quinolizin-3-yl)oxy)carbonyl)amino)ethyl)-1,2,-phenylenebis(2-ethylbutanoate)acetate (Compound-7),1-(((2-(4-amino-2,5-dioxo-2,3,4,5-tetrahydrobenzo[b][1,4]dioxocin-8-yl)ethyl)carbamoyl)oxy)ethylisobutyrate hydrofluoride (Compound-8a),1-(((2-(3-amino-2,5-dioxo-2,3,4,5-tetrahydrobenzo[b][1,4]dioxocin-8-yl)ethyl)carbamoyl)oxy)ethylisobutyrate hydrofluoride (Compound-8b),(5-(2-((ethoxycarbonyl)amino)ethyl)-2-(2-methylamino)acetoxy)phenylbenzoate hydrochloride (Compound-9a),5-(2-((ethoxycarbonyl)amino)ethyl)-2-(2-methylamino)acetoxy)phenylbenzoate hydrochloride (Compound-9b), 4-(2-aminoethyl)-1,2-phenylenedibenzoate hydrochloride (Compound-10),(S)-4-(2-amino-3-isopropoxy-3-oxopropyl)-1,2-phenylene dibenzoatehydrochloride (Compound-11),(S)-4-(2-amino-3-(heptan-4-yloxy)-3-oxopropyl)-1,2-phenylene dibenzoatehydrochloride (Compound-12),(S)-4-(2-amino-3-isopropoxy-3-oxopropyl)-1,2-phenylene dipentanoatehydrochloride (Compound-13),(S)-4-(2-amino-3-ethoxy-3-oxopropyl)-1,2-phenylene diacetatehydrochloride (Compound-14),(S)-4-(2-amino-3-oxo-3-(pentan-3-yloxy)propyl)-1,2-phenylenebis(2-methylpropanoate) hydrobromide (Compound-15),(S)-4-(2-aminoacetamido)-3-isopropoxy-3-oxopropyl)-1,2-phenylenedibenzoate hydrochloride (Compound-16),4-((2S)-3-oxo-3-(pentan-3-yloxy)-2-(pyrrolidine-2-carboxamido)propyl)-1,2-phenylenedibenzoate hydrofluoride (Compound-17),(S)-4-(3-isopropoxy-3-oxo-2-(piperidine-4-carboxamido)propyl)-1,2-phenylenedibenzoate hydrochloride (Compound-18),4-((2S)-3-isopropoxy-3-2-(octahydro-1H-quinolizine-2-carboxamido)-3-oxopropyl)-1,2-phenylenebis(2-methylpropanoate) hydrochloride (Compound-19), (2S)-isopropyl3-(3-amino-2,5-dioxo-2,3,4,5-tetrahydrobenzo[b][1,4]dioxocin-8-yl)-2-(((1-(isobutyryloxy)ethoxy)carbonyl)amino)propanoatehydrobromide (Compound-20a), (2S)-isopropyl3-(4-amino-2,5-dioxo-2,3,4,5-tetrahydrobenzo[b][1,4]dioxocin-8-yl)-2-(((1-(isobutyryloxy)ethoxy)carbonyl)amino)propanoatehydrobromide (Compound-20b),5-((2S)-2-(((1-(isobutyryloxy)ethoxy)carbonyl)amino-3-isopropoxy-3-oxopropyl)-2-(2-(methylamino)acetoxy)phenylbenzoate hydrochloride (Compound-21a),4-((2S)-2-(((1-(isobutyryloxy)ethoxy)carbonyl)amino-3-isopropoxy-3-oxopropyl)-2-(2-(methylamino)acetoxy)phenylbenzoate hydrochloride (Compound-21b),4-((2S)-3-isopropoxy-2-((((octahydroindolizin-1-yl)oxy)carbonyl)amino)-3-oxopropyl)-1,2-phenylenedibenzoate acetate (Compound-22), 2-(diethylamino)ethyl2-[(2,6-dichloro-3-methylphenyl)amino]benzoate.acetate (Compound-23),(Z)-2-(diethylaminoethyl)ethyl2-(5-fluoro-2-methyl-1-(4-methylsulfinyl)benzylidene)-1H-inden-1-yl)acetate.AcOH(Compound-24), 2-(dimethylamino)ethyl 2-(3-phenoxyphenyl)propionate.hydrochloride (Compound-25), S-(2-(dimethylamino)ethyl2-(3-phenoxyphenyl) propanethioate hydrochloride (Compound-26),2-(dipropylamino)ethyl4-acetoxy-2′,4′-difluoro-[1,1′-biphenyl]-3-carboxylate hydrochloride(Compound-27), 2-(diethylamino)ethyl 2-(4-isobutylphenyl)propionatehydrochloride (Compound-28), 2-(diethylamino)ethyl 2-acetoxybenzoatehydrochloride (Compound-29), 4-(2-aminoethyl)benzene-1,2-diolhydrochloride (Compound 30), 2-amino-3-(3,4-dihydroxyphenyl)propanoicacid (L-Dopa, Compound 31), acetylsalicylic acid (Compound 32),2-(ρ-isobutylphenyl)propionic acid (ibuprofen, Compound 33), and2-(3-phenoxyphenyl) propionic acid (Compound 34) through human skin weremeasured in vitro by using modified Franz cells, which were isolatedfrom human skin tissue (360-400 μm thick) of the anterior and posteriorthigh areas. The receiving fluid consisted of 10 mL of pH 7.4 phosphatebuffer (0.2 M) are shown in Table 1. The results suggested that thepositive charge on the amino group has a very important role in thepassage of the drug across the membrane and skin barrier.

TABLE 1 The Cumulative amounts of Pro-drugs of dopamine, L-Dopa, andNSAIDs in a period of 8 hours. The cumulative amount of test compounds 1h 2 h 4 h 8 h Compound-1 0.67 mg 1.33 mg 2.67 mg 5.25 mg Compound-2 0.65mg 1.29 mg 2.63 mg 5.23 mg Compound-3 0.61 mg 1.23 mg 2.49 mg 5.01 mgCompound-4 0.59 mg 1.21 mg 2.45 mg 4.93 mg Compound-5 0.60 mg 1.19 mg2.37 mg 4.79 mg Compound-6 0.63 mg 1.25 mg 2.53 mg 5.03 mg Compound-70.64 mg 1.27 mg 2.57 mg 5.05 mg Compound-8a 0.63 mg 1.22 mg 2.43 mg 4.83mg Compound-8b 0.62 mg 1.24 mg 2.47 mg 4.85 mg Compound-9a 0.61 mg 1.23mg 2.45 mg 4.87 mg Compound-9b 0.62 mg 1.26 mg 2.48 mg 4.95 mgCompound-10 0.68 mg 1.35 mg 2.74 mg 5.55 mg Compound-11 0.58 mg 1.15 mg2.33 mg 4.71 mg Compound-12 0.53 mg 1.07 mg 2.13 mg 4.29 mg Compound-130.52 mg 1.05 mg 2.10 mg 4.23 mg Compound-14 0.60 mg 1.21 mg 2.44 mg 4.82mg Compound-15 0.55 mg 1.08 mg 2.23 mg 4.41 mg Compound-16 0.56 mg 1.13mg 2.23 mg 4.49 mg Compound-17 0.58 mg 1.15 mg 2.33 mg 4.71 mgCompound-18 0.52 mg 1.03 mg 2.10 mg 4.19 mg Compound-19 0.48 mg 0.98 mg2.02 mg 4.09 mg Compound-20(a) 0.42 mg 0.88 mg 1.75 mg 3.49 mgCompound-20(b) 0.43 mg 0.85 mg 1.78 mg 3.52 mg Compound-21(a) 0.52 mg1.08 mg 2.17 mg 4.37 mg Compound-21(b) 0.51 mg 1.05 mg 2.08 mg 4.19 mgCompound-22 0.45 mg 0.89 mg 1.78 mg 3.61 mg Compound-23 0.85 mg 1.69 mg3.38 mg 6.85 mg Compound-24 0.87 mg 1.79 mg 3.53 mg 7.15 mg Compound-251.15 mg 2.31 mg 4.58 mg 9.22 mg Compound-26 1.17 mg 2.33 mg 4.65 mg 9.31mg Compound-27 1.01 mg 2.03 mg 4.11 mg 8.25 mg Compound-28 1.15 mg 2.28mg 4.57 mg 9.19 mg Compound-29 1.08 mg 2.13 mg 4.27 mg 8.48 mgCompound-30 0.12 mg 0.25 mg 0.52 mg 1.05 mg Compound-31 0.001 mg  0.001mg  0.002 mg  0.003 mg  Compound-32 0.001 mg  0.001 mg  0.002 mg  0.003mg  Compound-33 0.001 mg  0.001 mg  0.002 mg  0.003 mg  Compound-340.001 mg  0.001 mg  0.002 mg  0.003 mg 

Example 31 The Efficacy of(S)-4-(2-amino-3-isopropoxy-3-oxopropyl)-1,2-phenylene dibenzoatehydrochloride (Drug A) and 2-(diethylamino)ethyl 2-acetoxybenzoatehydrochloride (Drug B), on Improvement of Motor Function Deficits andReduction of Nigro-Striatal Neurodegeneration Induced by 6-OHDA inParkinson's Disease (PD) Model at HDB

Test subjects (Sprague-Dawley rats) were prepared, grouped, and testedfollowing the protocol below:

-   1. 200 Sprague-Dawley rats (male, 200-230 g) underwent brain    stereotaxic surgery after 1 week acclamation in animal facility;-   2. After anesthesia, a bur hole (1 mm in diameter) was made on skull    on the left side with a motor drill based on the coordinates: AP+0.5    mm, ML −2.8 mm, DV 6.0 mm relative to Bregma;-   3. 6-OHDA (20 μg at 5 mg/mL) or sterile saline was injected into the    left striatum with a microsyringe fitted with a 26-gauge steel    cannula. The chemical caused the retrograde neurodegeneration in the    distal substantia nigra and impairs dopamine transmission in the    nigro-striatal pathway, which eventually led to functional    disturbance of motor activities;-   4. Apomorphine and classic rotation test was carried out to screen    animals at the end of week 3 post 6-OHDA treatment. Animals that    matched lesion type and apomorphine-induced rotational scores were    allotted to 11 groups; and-   5. Rats were divided into 11 groups (n=12), each group was applied    with the drug(s) at the doses specified in Table 2:

TABLE 2 Doses and Drug(s) Applied to the Test Animals Group GroupDrug(s) Parent Dosage Administra- No. Name Applied drug (mg/kg) tionMethod 1 Positive L-DOPA N/A 6 Oral Control 2 Negative 30% N/A 1286Transdermal Control/ ethanol(v/v) (μL/kg) Vehicle (Vehicle) 3 Low doseDrug A L-DOPA 0.67 Transdermal 4 Moderate 2 Transdermal dose 5 High dose6 Transdermal 6 N/A Drug A L-DOPA 0.67 Transdermal Drug B Aspirin 30Transdermal 7 N/A Drug A L-DOPA 2 Transdermal Drug B Aspirin 30Transdermal 8 N/A Drug B Aspirin 30 Transdermal 9 N/A Drug A L-DOPA 0.67Transdermal Drug B Aspirin 90 Transdermal 10 N/A Drug A L-DOPA 2Transdermal Drug B Aspirin 90 Transdermal 11 N/A Drug B Aspirin 90Transdermal

-   6. Dose Formulations    -   (1) L-DOPA in water (3 mg/mL) was the positive control solution        for group 1 (orally for group 1, transdermally for groups 2-11).        The volume of administration was 2 mL/kg. Vehicle solution        (negative control solution for group 2) was 30% ethanol (v/v).        The volume of administration was 1286 μL/kg. Other test        solutions (groups 3-11) were freshly prepared every day.    -   (2) Preparation method of test solutions for groups 3-5: 50.33        mg of Drug A was dissolved in 10 mL of 30% ethanol (v/v). This        solution was the stock solution[(2)a].    -   a. The test solution for group 5 (high dose group) of Drug A:        The stock solution [(2)a] was the test solution for Group 5. The        volume of administration was 1286 μL/kg.    -   b. The test solution for group 4 (moderate dose group) of Drug        A: 3.00 mL of the stock solution [(2)a] was diluted to final        volume 9.00 mL with 30% ethanol (v/v). This solution was the        test solution for Group 4. The volume of administration was 1286        μL/kg.    -   c. The test solution for group 3 (low dose group) of Drug A:        1.00 mL of the stock solution [(2)a]) was diluted to final        volume 9.00 mL with 30% ethanol (v/v). This solution was the        test solution for Group 4. The volume of administration was 1286        μL/kg.    -   (3) Preparation method of test solutions for group 6: 5.67 mg of        Drug A and 264 mg of Drug B were dissolved in 10 mL of 30%        ethanol (v/v). This solution was the test solution for groups 6.    -   (4) Preparation method of test solutions for groups 7: 16.67 mg        of Drug A and 264 mg of Drug B were dissolved in 10 mL of 30%        ethanol (v/v). This solution was the test solution for groups 7.    -   (5) Preparation method of test solutions for groups 8: 264 mg of        Drug B was dissolved in 10 mL of 30% ethanol (v/v). This        solution was the test solution for groups 8.    -   (6) Preparation method of test solutions for group 9: 5.67 mg of        Drug A and 791 mg of Drug B were dissolved in 10 mL of 30%        ethanol (v/v). This solution was the test solution for groups 9.    -   (7) Preparation method of test solutions for groups 10: 16.67 mg        of Drug A and 791 mg of Drug B were dissolved in 10 mL of 30%        ethanol (v/v). This solution was the test solution for groups        10.    -   (8) Preparation method of test solutions for groups 11: 791 mg        of Drug A was dissolved in 10 mL of 30% ethanol (v/v). This        solution was the test solution for groups 11.-   7. On the day prior to the scheduled dosing, the hair (all rat    including the positive control group rats with L-DOPA due to the    double-blinded fashion) was removed from the up back (around the    neck and shoulder) of the animal using a small animal clipper. The    animals were re-clipped as necessary throughout the course of the    study to keep the epidermis exposed. On the day of dosing, an    aliquot (643 μL/kg) of the dosing formulation was administered to a    3 cm by 3 cm square section of the animal (around the neck and    shoulder) at around 9 am and repeat it at around 4 μm.-   8. At week 4 post lesion, the groups started to receive treatment.-   9. Rotarod tests were performed every week after treatment.

FIG. 1 showed the staying time results in the Totarod test describedabove after 1-3 week treatment (n=12). FIG. 2 showed the falling speedresults in Totarod test described above after 1-3 week treatment (n=12).

The improvement of performance in PD animals were supposed to staylonger on the road and endure a higher speed of the rotating (if no sideeffect of the test drug(s) was involved).

By pooling all batches data together (12 animals/group), the positivegroup (Group 1, L-DOPA, 6 mg/kg, orally) did not exhibit efficacycomparing to Vehicle group after three-week treatment. However, all testdrug treatment groups (Groups 2˜11) exhibited stronger efficacycomparing to Vehicle group after three-week treatment. Levodopacontributed to the development of motor complications in PD. Levodopacaused nausea, vomiting, gastrointestinal bleeding, dyskinesia at peakdose, and end-of-dose deterioration of function, so the positive group(Group 1, L-DOPA, 6 mg/kg, orally) did not exhibit efficacy comparing toVehicle group. However, the transdermally administrated pro-drug ofL-Dopa (Drug A) avoided dyskinesia at peak dose, and end-of-dosedeterioration of function. The efficacy of the low dose of Drug A (apro-drug of L-Dopa, 0.67 mg/kg, group 3) was much higher than that ofthe moderate and high dose groups of Drug A (2 mg and 6 mg/kg, groups 4and 5). Such result showed that the dose of transdermal administratedpro-drug of L-dopa was much less than that of the oral administratedL-dopa (9 times less). 30 mg and 90 mg/kg of Drug B (a pro-drug ofaspirin) exhibited good efficacy comparing to vehicle group and positivecontrol group after three-week treatment. 30 mg of Drug B (group 8)exhibited similar efficacy as 90 mg of Drug B (group 11). This resultshowed that 30 mg/kg dose might have been sufficient for Drug B andhigher dose was not necessary. Applying a combination of Drug A and DrugB (group 6 and group 9) to the subjects worked much better than applyingeither Drug B (groups 8 and 11) or Drug A (group 3) alone.

When applying a combination of a plurality of drugs (e.g. one or moreHPPs and/or other drug(s)) to a subject, each drug could be appliedseparately, or one or more of the drugs could be applied atsubstantially the same time as separate drugs (e.g. spraying two or moredrugs at substantially the same time without mixing the drugs beforespraying), or one or more drugs could be mixed together before applyingto the subject, or any combination of the above application methods. Thedrugs could be applied in any order possible.

Example 32 The Efficacy of Drug A (Transdermally), Drug B(Transdermally), and Carbidopa Orally on Improvement of Motor FunctionDeficits and Reduction of Nigro-Striatal Neurodegeneration Induced by6-OHDA in Parkinson's Disease (PD) Model at HDB

Test subjects (Sprague-Dawley rats) were prepared as described inExample 31, and grouped and tested following the protocol below:

-   1. Rats were divided into 11 groups (n=12), each group was applied    with the drug(s) at the doses specified in Table 3:

TABLE 3 Doses and Drug(s) Applied to the Test Animals Group GroupDrug(s) Parent Dosage Administra- No. Name Applied drug (mg/kg) tionMethod 1 Positive L-DOPA N/A 6 Oral Control Carbidopa N/A 1.5 Oral 2Negative 30% N/A 1,286 Transdermal Control ethanol(v/v) (μL/kg)(Vehicle) Carbidopa N/A 1.5 Oral 3 Low dose Drug A L-DOPA 0.67Transdermal Carbidopa N/A 1.5 Oral 4 Moderate Drug A L-DOPA 2Transdermal dose Carbidopa N/A 1.5 Oral 5 High dose Drug A L-DOPA 6Transdermal Carbidopa N/A 1.5 Oral 6 Drug A L-DOPA 0.67 Transdermal DrugB Aspirin 30 Transdermal Carbidopa N/A 1.5 Oral 7 Drug A L-DOPA 2Transdermal Drug B Aspirin 30 Transdermal Carbidopa N/A 1.5 Oral 8 DrugB Aspirin 30 Transdermal Carbidopa N/A 1.5 Oral 9 Drug A L-DOPA 0.67Transdermal Drug B Aspirin 90 Transdermal Carbidopa N/A 1.5 Oral 10 DrugA L-DOPA 2 Transdermal Drug B Aspirin 90 Transdermal Carbidopa N/A 1.5Oral 11 Drug B Aspirin 90 Transdermal Carbidopa N/A 1.5 Oral

-   2. Dose Formulations    -   (1) Both L-DOPA (3 mg/ml) and carbidopa (1.5 mg/mL) in water        were used as the positive control solution for group 1 (orally),        the volume of administration was 2 mL/kg. Carbidopa in water (3        mg/mL) was as an aromatic-L-amino-acid decarboxylase inhibitor        for groups 2-11 (orally), the volume of administration was 2        mL/kg. Vehicle solution (negative control solution for group 2)        was 30% ethanol (v/v), the volume of administration was 1,286        μL/kg. Other test solutions (transdermally, groups 3-11) were        freshly prepared every day.    -   (2) Preparation method of test solutions for groups 3-5: 50.33        mg of Drug A was dissolved in 10 mL of 30% ethanol (v/v). This        solution was the stock solution (2).    -   (a) The stock solution (2) was the test solution for Group 5        (high dose group) of Drug A. The volume of administration was        1,286 μL/kg;    -   (b) The test solution for group 4 (moderate dose group) of Drug        A: 3.00 mL of the stock solution (2) was diluted to final volume        9.00 mL with 30% ethanol (v/v). This solution was the test        solution for Group 4. The volume of administration was 1,286        μL/kg;    -   (c) The test solution for group 3 (low dose group) of Drug A:        1.00 mL of the stock solution (2) was diluted to final volume        9.00 mL with 30% ethanol (v/v). This solution was the test        solution for Group 3. The volume of administration was 1,286        μL/kg.    -   (3) Preparation method of test solutions for group 6: 5.67 mg of        Drug A and 264 mg of Drug B were dissolved in 10 mL of 30%        ethanol (v/v). This solution was the test solution for groups 6;    -   (4) Preparation method of test solutions for groups 7: 16.67 mg        of Drug A and 264 mg of Drug B were dissolved in 10 mL of 30%        ethanol (v/v). This solution was the test solution for groups 7;    -   (5) Preparation method of test solutions for groups 8: 264 mg of        Drug B was dissolved in 10 mL of 30% ethanol (v/v). This        solution was the test solution for groups 8;    -   (6) Preparation method of test solutions for group 9: 5.67 mg of        Drug A and 791 mg of Drug B were dissolved in 10 mL of 30%        ethanol (v/v). This solution was the test solution for groups 9;    -   (7) Preparation method of test solutions for groups 10: 16.67 mg        of Drug A and 791 mg of Drug B were dissolved in 10 mL of 30%        ethanol (v/v). This solution was the test solution for groups        10;    -   (8) Preparation method of test solutions for groups 11: 791 mg        of Drug B was dissolved in 10 mL of 30% ethanol (v/v). This        solution was the test solution for groups 11.-   3. On the day prior to the scheduled dosing, the hair (all rat    including the positive control group rats with L-DOPA due to the    double-blinded fashion) was removed from the up back (around the    neck and shoulder) of the animal using a small animal clipper. The    animals were re-clipped as necessary throughout the course of the    study to keep the epidermis exposed. On the day of dosing, an    aliquot (643 μL/kg) of the dosing formulation was administered to a    3 cm by 3 cm square section of the animal (around the neck and    shoulder) at around 9 am and repeat it at around 4 μm.-   4. At week 4 post lesion, the groups started to receive treatment.-   5. Rotarod test were performed 4 weeks after treatment.

FIG. 3 showed the staying time results in Totarod test described aboveafter 4 weeks treatment (n=12). FIG. 4 showed the falling speed resultsin Totarod test described above after 4 weeks treatment (n=12).

Carbidopa can decrease peripheral DDC conversion of levodopa before itcrosses the blood-brain barrier, then reduced the side effects ofL-Dopa. The results show that carbidopa increased efficacy of all L-Dopaand the pro-drug of L-Dopa (Drug A) treated groups (groups 1, 3, 4, 5,6, 7, 9, 10), but did not change the efficacy of vehicle group andpro-drug of aspirin treated groups (Drug B) (groups 2, 8 and 11).

When applying a combination of a plurality of drugs (e.g. one or moreHPPs and/or other drug(s)) to a subject, each drug could be appliedseparately, or one or more of the drugs could be applied atsubstantially the same time as separate drugs (e.g. spraying two or moredrugs at substantially the same time without mixing the drugs beforespraying), or one or more drugs could be mixed together before applyingto the subject, or any combination of the above application methods. Thedrugs could be applied in any order possible.

Example 33 The Efficacy of Pro-Drugs of L-Dopa and Ibuprofen onImprovement of Motor Function Deficits and Reduction of Nigro-StriatalNeurodegeneration Induced by 6-OHDA in Parkinson's Disease (PD) Model atHDB

Test subjects (Sprague-Dawley rats) were prepared as described inExample 31, and grouped and tested following the protocol below:

-   1. Rats were divided into 11 groups (n=12), each group was applied    with the drug(s) at the doses specified in Table 4, Drug C was    (S)-4-(2-amino-3-oxo-3-(pentan-3-yloxy)propyl)-1,2-phenylene    bis(2-methylpropanoate) hydrobromide, and Drug D was    2-(diethylamino)ethyl 2-(4-isobutylphenyl)propionate acetic acid    salt:

TABLE 4 Doses and Drug(s) Applied to the Test Animals Group GroupDrug(s) Parent Dosage Administra- No. Name Applied drug (mg/kg) tionMethod 1 Positive L-DOPA N/A 6 Oral Control 2 Negative 30% N/A 1286Transdermal Control ethanol(v/v) (μL/kg) (Vehicle) 3 Low dose Drug CL-DOPA 0.67 Transdermal 4 Moderate 2 Transdermal dose 5 High dose 6Transdermal 6 Drug C L-DOPA 0.67 Transdermal Drug D Ibuprofen 15Transdermal 7 Drug C L-DOPA 2 Transdermal Drug D Ibuprofen 15Transdermal 8 Drug D Ibuprofen 15 Transdermal 9 Drug C L-DOPA 0.67Transdermal Drug D Ibuprofen 45 Transdermal 10 Drug C L-DOPA 2Transdermal Drug D Ibuprofen 45 Transdermal 11 Drug D Ibuprofen 45Transdermal

-   2. Dose Formulations-   (1) L-DOPA in water (3 mg/mL) was the positive control solution for    group 1 (orally), the volume of administration was 2 mL/kg. Vehicle    solution (negative control solution for group 2) was 30% ethanol    (v/v), the volume of administration was 1286 μL/kg. Other test    solutions (groups 3-11) were freshly prepared every day.-   (2) Preparation method of test solutions for groups 3-5: 50.33 mg of    Drug C was dissolved in 10 mL of 30% ethanol (v/v). This solution    was the stock solution (2).    -   a) The test solution for group 5 (high dose group) of Drug C:        The stock solution (2) was the test solution for group 5. The        volume of administration was 1,286 μL/kg;    -   b) The test solution for group 4 (moderate dose group) of Drug        C: 3.00 mL of the stock solution (2) was diluted to final volume        9.00 mL with 30% ethanol (v/v). This solution was the test        solution for group 4. The volume of administration was 1,286        μL/kg;    -   c) The test solution for group 3 (low dose group) of Drug C:        1.00 mL of the stock solution (2) was diluted to final volume        9.00 mL with 30% ethanol (v/v). This solution was the test        solution for group 3. The volume of administration was 1,286        μL/kg.-   (3) Preparation method of test solutions for group 6: 5.67 mg of    Drug C and 132 mg of Drug D were dissolved in 10 mL of 30% ethanol    (v/v). This solution was the test solution for group 6.-   (4) Preparation method of test solutions for groups 7: 16.67 mg of    Drug C and 132 mg of Drug D were dissolved in 10 mL of 30% ethanol    (v/v). This solution was the test solution for group 7.-   (5) Preparation method of test solutions for groups 8: 132 mg of    Drug D was dissolved in 10 mL of 30% ethanol (v/v). This solution    was the test solution for group 8.-   (6) Preparation method of test solutions for group 9: 5.67 mg of    Drug C and 395.5 mg of Drug D were dissolved in 10 mL of 30% ethanol    (v/v). This solution was the test solution for group 9.-   (7) Preparation method of test solutions for groups 10: 16.67 mg of    Drug C and 395.5 mg of Drug D were dissolved in 10 mL of 30% ethanol    (v/v). This solution was the test solution for group 10.-   (8) Preparation method of test solutions for groups 11: 395.5 mg of    Drug D was dissolved in 10 mL of 30% ethanol (v/v). This solution    was the test solution for groups 11.-   3. On the day prior to the scheduled dosing, the hair (all rat    including the positive control group rats with L-DOPA due to the    double-blinded fashion) was removed from the up back (around the    neck and shoulder) of the animal using a small animal clipper. The    animals were re-clipped as necessary throughout the course of the    study to keep the epidermis exposed. On the day of dosing, an    aliquot (643 μL/kg) of the dosing formulation was administered to a    3 cm by 3 cm square section of the animal (around the neck and    shoulder) at around 9 am and repeat it at around 4 μm.-   4. At week 4 post lesion, the groups started to receive treatment.-   5. Rotarod test will be performed 4 weeks after treatment.

FIG. 5 showed the staying time results in Totarod test described aboveafter 4 weeks treatment (n=12). FIG. 6 showed the falling speed resultsin Totarod test described above after 4 weeks treatment (n=12).

By pooling all batches data together (12 animals/group), the positivegroup (L-DOPA, 6 mg/kg, orally) did not exhibit efficacy comparing tothe negative control group after four-week treatment, however, all testdrug(s) treated groups exhibited stronger efficacy comparing to vehiclegroup after four-week treatment. Levodopa caused nausea, vomiting,gastrointestinal bleeding, dyskinesia at peak dose, and end-of-dosedeterioration of function, so the positive group (L-DOPA, 6 mg/kg,orally) did not exhibit efficacy comparing to Vehicle group. However,the transdermally administrated pro-drug of L-Dopa (Compound C) avoideddyskinesia at peak dose, and end-of-dose deterioration of function. Theefficacy of the low dose of Drug C (the pro-drug of L-Dopa, 0.67 mg/kg,group 3) was much higher than that of the moderate and high dose groupsof Drug C (2 mg and 6 mg/kg, groups 4 and 5) and this result showed thatthe dose of transdermally administrated pro-drug of L-dopa was much lessthan that of orally administrated L-dopa. 15 mg and 45 mg/kg of Drug D(the pro-drug of ibuprofen) exhibited good efficacy comparing to vehiclegroup and positive control group after four-week treatment. 15 mg ofDrug D (group 8) exhibited similar efficacy as 45 mg of Drug D (group11), so 15 mg/kg dose was enough and higher dose was not necessary.Applying a combination of Drug C and Drug D (group 6 and group 9) workedmuch better than applying either Drug D (group 8, and 11) or Drug C(group 3) alone.

Example 34 The Efficacy of Pro-Drugs of L-Dopa and Ibuprofen and 1.5Mg/Kg of Carbidopa on Improvement of Motor Function Deficits andReduction of Nigro-Striatal Neurodegeneration Induced by 6-OHDA inParkinson's Disease (PD) Model at HDB

Test subjects (Sprague-Dawley rats) were prepared as described inExample 31, and grouped and tested following the protocol below:

-   1. Rats were divided into 11 groups (n=12), each group was applied    with the drug(s) at the doses specified in Table 5:

TABLE 5 Doses and Drug(s) Applied to the Test Animals Group GroupDrug(s) Parent Dosage Administra- No. Name Applied drug (mg/kg) tionMethod 1 Positive L-DOPA N/A 6 Oral Control Carbidopa N/A 1.5 Oral 2Negative 30% N/A 1,286 Transdermal Control ethanol(v/v) (μL/kg)(Vehicle) Carbidopa N/A 1.5 Oral 3 Low dose Drug C L-DOPA 0.67Transdermal Carbidopa N/A 1.5 Oral 4 Moderate Drug C L-DOPA 2Transdermal dose Carbidopa N/A 1.5 Oral 5 High dose Drug C L-DOPA 6Transdermal Carbidopa N/A 1.5 Oral 6 Drug C L-DOPA 0.67 Transdermal DrugD Ibuprofen 15 Transdermal Carbidopa N/A 1.5 Oral 7 Drug C L-DOPA 2Transdermal Drug D Ibuprofen 15 Transdermal Carbidopa N/A 1.5 Oral 8Drug D Ibuprofen 15 Transdermal Carbidopa N/A 1.5 Oral 9 Drug C L-DOPA0.67 Transdermal Drug D Ibuprofen 45 Transdermal Carbidopa N/A 1.5 Oral10 Drug C L-DOPA 2 Transdermal Drug D Ibuprofen 45 Transdermal CarbidopaN/A 1.5 Oral 11 Drug D Ibuprofen 45 Transdermal Carbidopa N/A 1.5 Oral

-   2. Dose Formulations-   (1) Both L-DOPA (3 mg/ml) and carbidopa (1.5 mg/mL) in water was    used as the positive control solution for group 1 (orally). The    volume of administration was 2 mL/kg. Carbidopa in water (1.5 mg/mL)    as an aromatic-L-amino-acid decarboxylase inhibitor was applied to    groups 2-11 (orally), the volume of administration was 2 mL/kg.    Vehicle solution (negative control solution for group 2) was 30%    ethanol (v/v), the volume of administration was 1,286 μL/kg. Other    test solutions (transdermally, groups 3-11) were freshly prepared    every day.-   (2) Preparation method of test solutions for groups 3-5: 50.33 mg of    Drug C was dissolved in 10 mL of 30% ethanol (v/v). This solution    was the stock solution (2).    -   a. Preparation of Drug C solution as the test solution for group        5 (high dose group). The stock solution (2) was the test        solution for Group 5. The volume of administration was 1,286        μL/kg;    -   b. Preparation of Drug C solution as the test solution for group        4 (moderate dose group): 3.00 mL of the stock solution (2) was        diluted to final volume 9.00 mL with 30% ethanol (v/v). This        solution was the test solution for Group 4. The volume of        administration was 1,286 μL/kg;    -   c. Preparation of Drug C solution as the test solution for group        3 (low dose group): 1.00 mL of the stock solution (2) was        diluted to final volume 9.00 mL with 30% ethanol (v/v). This        solution was the test solution for Group 4. The volume of        administration was 1,286 μL/kg.-   (3) Preparation method of test solutions for group 6: 5.67 mg of    Drug C and 132 mg of Drug D were dissolved in 10 mL of 30% ethanol    (v/v). This solution was the test solution for groups 6.-   (4) Preparation method of test solutions for groups 7: 16.67 mg of    Drug C and 132 mg of Drug D were dissolved in 10 mL of 30% ethanol    (v/v). This solution was the test solution for groups 7.-   (5) Preparation method of test solutions for groups 8: 132 mg of    Drug D was dissolved in 10 mL of 30% ethanol (v/v). This solution    was the test solution for groups 8.-   (6) Preparation method of test solutions for group 9: 5.67 mg of    Drug C and 395.5 mg of Drug D were dissolved in 10 mL of 30% ethanol    (v/v). This solution was the test solution for groups 9.-   (7) Preparation method of test solutions for groups 10: 16.67 mg of    Drug C and 395.5 mg of Drug D were dissolved in 10 mL of 30% ethanol    (v/v). This solution was the test solution for groups 10.-   (8) Preparation method of test solutions for groups 11: 395.5 mg of    Drug D was dissolved in 10 mL of 30% ethanol (v/v). This solution    was the test solution for groups 11.-   3. On the day prior to the scheduled dosing, the hair (all rat    including the positive control group rats with L-DOPA due to the    double-blinded fashion) was removed from the up back (around the    neck and shoulder) of the animal using a small animal clipper. The    animals were re-clipped as necessary throughout the course of the    study to keep the epidermis exposed. On the day of dosing, an    aliquot (643 μL/kg) of the dosing formulation was administered to a    3 cm by 3 cm square section of the animal (around the neck and    shoulder) at around 9 am and repeat it at around 4 pm.-   4. At week 4 post lesion, the groups started to receive treatment.-   5. Rotarod test was performed 4 weeks after treatment.

FIG. 7 showed the staying time results in Totarod test described aboveafter 4 weeks treatment (n=12). FIG. 8 showed the falling speed resultsin Totarod test described above after 4 weeks treatment (n=12).

Carbidopa can decrease peripheral DDC conversion of levodopa before itcrosses the blood-brain barrier, then reduced the side effects ofL-Dopa. The results show that carbidopa increased efficacy of all L-Dopaand the pro-drug of L-Dopa treated groups (groups 1, 3, 4, 5, 6, 7, 9,10), but did not change the efficacy of vehicle group and pro-drug ofibuprofen treated groups (groups 2, 8 and 11).

Example 35 The Efficacy of4-(2-(((1-((pyrrolidine-2-carbonyl)oxy)ethoxy)amino)ethyl)-1,2,-phenylenedibenzoate hydrochloride (Drug E) and 4-(dimethylamino)butyl2-(3-phenoxyphenyl) propionate hydrochloride (Drug F) on Improvement ofMotor Function Deficits and Reduction of Nigro-StriatalNeurodegeneration Induced by 6-OHDA in Parkinson's Disease (PD) Model atHDB

Test subjects (Sprague-Dawley rats) were prepared as described inExample 31, and grouped and tested following the protocol below:

-   1. Rats were divided into 11 groups (n=12), each group was applied    with the drug(s) at the doses specified in Table 6:

TABLE 6 Doses and Drug(s) Applied to the Test Animals Group GroupDrug(s) Parent Dosage Administra- No. Name Applied drug (mg/kg) tionMethod 1 Positive L-DOPA N/A 6 Oral Control 2 Negative 30% N/A 1,286Transdermal Control ethanol(v/v) (μL/kg) (Vehicle) 3 Low dose Drug EDopamine 0.5 Transdermal 4 Moderate 1.5 Transdermal dose 5 High dose 4.5Transdermal 6 Drug E Dopamine 0.5 Transdermal Drug F Fenoprofen 20Transdermal 7 Drug E Dopamine 1.5 Transdermal Drug F Fenoprofen 20Transdermal 8 Drug F Fenoprofen 20 Transdermal 9 Drug E Dopamine 0.5Transdermal Drug F Fenoprofen 60 Transdermal 10 Drug E Dopamine 1.5Transdermal Drug F Fenoprofen 60 Transdermal 11 Drug F Fenoprofen 60Transdermal

-   2. Dose Formulations-   (1) L-DOPA in water (3 mg/mL) was the positive control solution for    group 1 (orally for group 1). The volume of administration was 2    mL/kg. Vehicle solution (negative control solution for group 2) was    30% ethanol (v/v). The volume of administration was 1286 μL/kg.    Other test solutions (groups 3-11) were freshly prepared every day.-   (2) Preparation method of test solutions for groups 3-5: 37.75 mg of    Drug E was dissolved in 10 mL of 30% ethanol (v/v). This solution    was the stock solution (2).    -   a. The test solution for group 5 (high dose group) of Drug E:        The stock solution (2) was the test solution for Group 5. The        volume of administration was 1,286 μL/kg;    -   b. The test solution for group 4 (moderate dose group) of Drug        E: 3.00 mL of the stock solution (2) was diluted to final volume        9.00 mL with 30% ethanol (v/v). This solution was the test        solution for Group 4. The volume of administration was 1286        μL/kg;    -   c. The test solution for group 3 (low dose group) of Drug E:        1.00 mL of the stock solution (2) was diluted to final volume        9.00 mL with 30% ethanol (v/v). This solution was the test        solution for Group 3. The volume of administration was 1286        μL/kg.-   (3) Preparation method of test solutions for group 6: 4.25 mg of    Drug E and 176 mg of Drug E were dissolved in 10 mL of 30% ethanol    (v/v). This solution was the test solution for groups 6.-   (4) Preparation method of test solutions for groups 7: 12.5 mg of    Drug E and 176 mg of Drug E were dissolved in 10 mL of 30% ethanol    (v/v). This solution was the test solution for groups 7.-   (5) Preparation method of test solutions for groups 8: 176 mg of    Drug E was dissolved in 10 mL of 30% ethanol (v/v). This solution    was the test solution for groups 8.-   (6) Preparation method of test solutions for group 9: 4.25 mg of    (Drug E and 527 mg of Drug E were dissolved in 10 mL of 30% ethanol    (v/v). This solution was the test solution for groups 9.-   (7) Preparation method of test solutions for groups 10: 12.5 mg of    Drug E and 527 mg of Drug E were dissolved in 10 mL of 30% ethanol    (v/v). This solution was the test solution for groups 10.-   (8) Preparation method of test solutions for groups 11: 527 mg of 4    Drug E was dissolved in 10 mL of 30% ethanol (v/v). This solution    was the test solution for groups 11.-   3. On the day prior to the scheduled dosing, the hair (all rat    including the positive control group rats with L-DOPA due to the    double-blinded fashion) was removed from the up back (around the    neck and shoulder) of the animal using a small animal clipper. The    animals were re-clipped as necessary throughout the course of the    study to keep the epidermis exposed. On the day of dosing, an    aliquot (643 μL/kg) of the dosing formulation was administered to a    3 cm by 3 cm square section of the animal (around the neck and    shoulder) at around 9 am and repeat it at around 4 μm.-   4. At week 4 post lesion, the groups started to receive treatment.-   5. Rotarod test was performed 4 weeks after treatment.

FIG. 9 showed the staying time results in Totarod test described aboveafter 4 weeks treatment (n=12). FIG. 10 showed the falling speed resultsin Totarod test described above after 4 weeks treatment (n=12).

By pooling all batches data together (12 animals/group), the positivegroup (L-DOPA, 6 mg/kg, orally) did not exhibit efficacy comparing toVehicle group after four-week treatment. However, all test drug(s)treated groups exhibited stronger efficacy comparing to vehicle groupafter four-week treatment. Levodopa and dopamine caused nausea,vomiting, gastrointestinal bleeding, dyskinesia at peak dose, andend-of-dose deterioration of function, so the positive group (L-DOPA, 6mg/kg, orally) did not exhibit efficacy comparing to vehicle group.However, the transdermally administrated pro-drug of dopamine avoideddyskinesia at peak dose, and end-of-dose deterioration of function. Theefficacy of the low dose of Drug E (the pro-drug of dopamine, 0.5 mg/kg,group 3) was much higher than that of the moderate and high dose groupsof Drug E (1.5 mg and 4.5 mg/kg, group 4 and 5), so higher dose of DrugE may have caused more side effects as L-Dopa did. 20 mg and 60 mg/kg ofDrug F exhibited good efficacy comparing to vehicle group and positivecontrol group after four-week treatment. 20 mg/Kg of Drug F (group 8)exhibited similar efficacy as 60 mg/Kg of Drug F (group 11). This resultshowed that 20 mg/kg dose was sufficient and higher dose was notnecessary. Applying a combination of Drug E and Drug F (group 6 andgroup 9) worked much better than applying either Drug F (groups 8 and11) or (Drug E (group 3) alone.

When applying a combination of a plurality of drugs (e.g. one or moreHPPs and/or other drug(s)) to a subject, each drug could be appliedseparately, or one or more of the drugs could be applied atsubstantially the same time as separate drugs (e.g. spraying two or moredrugs at substantially the same time without mixing the drugs beforespraying), or one or more drugs could be mixed together before applyingto the subject, or any combination of the above application methods. Thedrugs could be applied in any order possible.

Example 36 The Efficacy of Drug G((S)-4-(2-amino-3-(heptan-4-yloxy)-3-oxopropyl)-1,2-phenylene dibenzoatehydrochloride) and Drug H (2-(dipropylamino)ethyl4-acetoxy-2′,4′-difluoro-[1,1′-biphenyl]-3-carboxylate hydrochloride) onImprovement of Motor Function Deficits and Reduction of Nigro-StriatalNeurodegeneration Induced by 6-OHDA in Parkinson's Disease (PD) Model atHDB

Test subjects (Sprague-Dawley rats) were prepared as described inExample 31, and grouped and tested following the protocol below:

-   1. Rats were divided into 11 groups (n=12), each group was applied    with the drug(s) at the doses specified in Table 7:

TABLE 7 Doses and Drug(s) Applied to the Test Animals Group GroupDrug(s) Parent Dosage Administra- No. Name Applied drug (mg/kg) tionMethod 1 Positive L-DOPA N/A 6 Oral Control 2 Negative 30% N/A 1,286Transdermal Control ethanol(v/v) (μL/kg) (Vehicle) 3 Low dose Drug GL-DOPA 0.67 Transdermal 4 Moderate 2 Transdermal dose 5 High dose 6Transdermal 6 Drug G L-DOPA 0.67 Transdermal Drug H Diflunisal 15Transdermal 7 Drug G L-DOPA 0.67 Transdermal Drug H Diflunisal 15Transdermal 8 Drug H Diflunisal 15 Transdermal 9 Drug G L-DOPA 0.67Transdermal Drug H Diflunisal 45 Transdermal 10 Drug G L-DOPA 2Transdermal Drug H Diflunisal 45 Transdermal 11 Drug H Diflunisal 45Transdermal

-   2. Dose Formulations-   (1) L-DOPA in water (3 mg/mL) was the positive control solution for    group 1 (orally), the volume of administration was 2 mL/kg. Vehicle    solution (negative control solution for group 2) was 30% ethanol    (v/v), the volume of administration was 1,286 μL/kg. Other test    solutions (groups 3-11) were freshly prepared every day.-   (2) Preparation method of test solutions for groups 3-5: 50.33 mg of    Drug G was dissolved in 10 mL of 30% ethanol (v/v). This solution    was the stock solution (2).    -   a. The test solution for group 5 (high dose group) of Drug G:        The stock solution (2) was the test solution for Group 5. The        volume of administration was 1286 μL/kg;    -   b. The test solution for group 4 (moderate dose group) of Drug        G: 3.00 mL of the stock solution (2) was diluted to final volume        9.00 mL with 30% ethanol (v/v). This solution was the test        solution for Group 4. The volume of administration was 1286        μL/kg.    -   c. The test solution for group 3 (low dose group) of Drug G:        1.00 mL of the stock solution (2) was diluted to final volume        9.00 mL with 30% ethanol (v/v). This solution was the test        solution for Group 4. The volume of administration was 1286        μL/kg-   (3) Preparation method of test solutions for group 6: 5.67 mg of    Drug G and 132 mg of Drug H were dissolved in 10 mL of 30% ethanol    (v/v). This solution was the test solution for groups 6.-   (4) Preparation method of test solutions for groups 7: 16.67 mg of    Drug G and 132 mg of Drug H were dissolved in 10 mL of 30% ethanol    (v/v). This solution was the test solution for groups 7.-   (5) Preparation method of test solutions for groups 8: 132 mg of    Drug H was dissolved in 10 mL of 30% ethanol (v/v). This solution    was the test solution for groups 8.-   (6) Preparation method of test solutions for group 9: 5.67 mg of    Drug G and 395.5 mg of Drug H were dissolved in 10 mL of 30% ethanol    (v/v). This solution was the test solution for groups 9.-   (7) Preparation method of test solutions for groups 10: 16.67 mg of    Drug G and 395.5 mg of Drug H were dissolved in 10 mL of 30% ethanol    (v/v). This solution was the test solution for groups 10.-   (8) Preparation method of test solutions for groups 11: 395.5 mg of    Drug H was dissolved in 10 mL of 30% ethanol (v/v). This solution    was the test solution for groups 11.-   3. On the day prior to the scheduled dosing, the hair (all rat    including the positive control group rats with L-DOPA due to the    double-blinded fashion) was removed from the up back (around the    neck and shoulder) of the animal using a small animal clipper. The    animals were re-clipped as necessary throughout the course of the    study to keep the epidermis exposed. On the day of dosing, an    aliquot (643 μL/kg) of the dosing formulation was administered to a    3 cm by 3 cm square section of the animal (around the neck and    shoulder) at around 9 am and repeat it at around 4 μm.-   4. At week 4 post lesion, the groups started to receive treatment.-   5. Rotarod test was performed four weeks after treatment.

FIG. 11 showed the staying time results in Totarod test described aboveafter 4 weeks treatment (n=12). FIG. 12 showed the falling speed resultsin Totarod test described above after 4 weeks treatment (n=12).

By pooling all batches data together (12 animals/group), the positivegroup (L-DOPA, 6 mg/kg, orally) did not exhibit efficacy comparing tovehicle group after four-week treatment. However, all test drug(s)treatment groups exhibited stronger efficacy comparing to vehicle groupafter four-week treatment. Levodopa caused nausea, vomiting,gastrointestinal bleeding, dyskinesia at peak dose, and end-of-dosedeterioration of function, so the positive group (L-DOPA, 6 mg/kg,orally) did not exhibit efficacy comparing to vehicle group. However,the transdermally administrated pro-drug of L-Dopa avoided dyskinesia atpeak dose, and end-of-dose deterioration of function. The efficacy ofthe low dose of Drug G (the pro-drug of L-Dopa, 0.67 mg/kg, group 3) wasmuch higher than that of the moderate and high dose groups of Drug G (2mg and 6 mg/kg, groups 4 and 5). The higher dose may have caused sideeffects as L-dopa did. 15 mg and 45 mg/kg of Drug H exhibited goodefficacy comparing to vehicle group and positive control group afterfour-week treatment. 15 mg of Drug H (group 8) exhibited similarefficacy as 45 mg of Drug H (group 11). This suggested 15 mg/kg dose wassufficient and higher dose was not necessary. Applying a combination ofDrug G and Drug H (group 6 and group 9) worked much better than applyingeither Drug H (groups 8 and 11) or Drug G (groups 3) alone.

When applying a combination of a plurality of drugs (e.g. one or moreHPPs and/or other drug(s)) to a subject, each drug could be appliedseparately, or one or more of the drugs could be applied atsubstantially the same time as separate drugs (e.g. spraying two or moredrugs at substantially the same time without mixing the drugs beforespraying), or one or more drugs could be mixed together before applyingto the subject, or any combination of the above application methods. Thedrugs could be applied in any order possible.

Example 37 The Efficacy of Drug A (Transdermally), Drug B(Transdermally), and Entacapone (Orally) on Improvement of MotorFunction Deficits and Reduction of Nigro-Striatal NeurodegenerationInduced by 6-OHDA in Parkinson's Disease (PD) Model at HDB

Test subjects (Sprague-Dawley rats) were prepared as described inExample 31, and grouped and tested following the protocol below:

-   1. Rats were divided into 11 groups (n=12), each group was applied    with the drug(s) at the doses specified in Table 8:

TABLE 8 Doses and Drug(s) Applied to the Test Animals Group GroupDrug(s) Parent Dosage Administra- No. Name Applied drug (mg/kg) tionMethod 1 Positive L-DOPA N/A 6 Oral Control Entacapone N/A 25 Oral 2Negative 30% N/A 1,286 Transdermal Control ethanol(v/v) (μL/kg)(Vehicle) Entacapone N/A 25 Oral 3 Low dose Drug A L-DOPA 0.67Transdermal Entacapone N/A 25 Oral 4 Moderate Drug A L-DOPA 2Transdermal dose Entacapone N/A 25 Oral 5 High dose Drug A L-DOPA 6Transdermal Entacapone N/A 25 Oral 6 Drug A L-DOPA 0.67 Transdermal DrugB Aspirin 30 Transdermal Entacapone N/A 25 Oral 7 Drug A L-DOPA 2Transdermal Drug B Aspirin 30 Transdermal Entacapone N/A 25 Oral 8 DrugB Aspirin 30 Transdermal Entacapone N/A 25 Oral 9 Drug A L-DOPA 0.67Transdermal Drug B Aspirin 90 Transdermal Entacapone N/A 25 Oral 10 DrugA L-DOPA 2 Transdermal Drug B Aspirin 90 Transdermal Entacapone N/A 25Oral 11 Drug B Aspirin 90 Transdermal Entacapone N/A 25 Oral

-   2. Dose Formulations-   (1) Both L-DOPA (3 mg/ml) and entacapone (12.5 mg/mL) in 0.5% CMC-Na    (carboxymethylcellulose sodium salt) was the positive control    solution for group 1 (orally), the volume of administration was 2    mL/kg. Entacapone (12.5 mg/ml) as a catechol-O-methyl transferase    inhibitor in 0.5% CMC-Na was applied to groups 2-11 (orally), the    volume of administration was 2 mL/kg. Vehicle solution (negative    control solution for group 2) was 30% ethanol (v/v), the volume of    administration was 1,286 μL/kg. Other test solutions (transdermally,    groups 3-11) were freshly prepared every day.-   (2) Preparation method of test solutions for groups 3-5: 50.33 mg of    Drug A was dissolved in 10 mL of 30% ethanol (v/v). This solution    was the stock solution (2).    -   a. The stock solution (2) was the test solution for Group 5        (high dose group) of Drug A. The volume of administration was        1,286 μL/kg;    -   b. The test solution for group 4 (moderate dose group) of Drug        A: 3.00 mL of the stock solution (2) was diluted to final volume        9.00 mL with 30% ethanol (v/v). This solution was the test        solution for Group 4. The volume of administration was 1,286        μL/kg;    -   c. The test solution for group 3 (low dose group) of Drug A:        1.00 mL of the stock solution (2) was diluted to final volume        9.00 mL with 30% ethanol (v/v). This solution was the test        solution for Group 4. The volume of administration was 1286        μL/kg.-   (3) Preparation method of test solutions for group 6: 5.67 mg of    Drug A and 264 mg of Drug B were dissolved in 10 mL of 30% ethanol    (v/v). This solution was the test solution for groups 6;-   (4) Preparation method of test solutions for groups 7: 16.67 mg of    Drug A and 264 mg of Drug B were dissolved in 10 mL of 30% ethanol    (v/v). This solution was the test solution for groups 7;-   (5) Preparation method of test solutions for groups 8: 264 mg of    Drug B was dissolved in 10 mL of 30% ethanol (v/v). This solution    was the test solution for groups 8;-   (6) Preparation method of test solutions for group 9: 5.67 mg of    Drug A and 791 mg of Drug B were dissolved in 10 mL of 30% ethanol    (v/v). This solution was the test solution for groups 9;-   (7) Preparation method of test solutions for groups 10: 16.67 mg of    Drug A and 791 mg of Drug B were dissolved in 10 mL of 30% ethanol    (v/v). This solution was the test solution for groups 10;-   (8) Preparation method of test solutions for groups 11: 791 mg of    Drug B was dissolved in 10 mL of 30% ethanol (v/v). This solution    was the test solution for groups 11.-   3. On the day prior to the scheduled dosing, the hair (all rat    including the positive control group rats with L-DOPA due to the    double-blinded fashion) was removed from the up back (around the    neck and shoulder) of the animal using a small animal clipper. The    animals were re-clipped as necessary throughout the course of the    study to keep the epidermis exposed. On the day of dosing, an    aliquot (643 μL/kg) of the dosing formulation was administered to a    3 cm by 3 cm square section of the animal (around the neck and    shoulder) at around 9 am and repeat it at around 4 m.-   4. At week 4 post lesion, the groups started to receive treatment.-   5. Rotarod test was performed 4 weeks after treatment.

FIG. 13 showed the staying time results in Totarod test described aboveafter 4 weeks treatment (n=12). FIG. 14 showed the falling speed resultsin Totarod test described above after 4 weeks treatment (n=12).

Entacapone was Catechol-O-methyl transferase inhibitors and preventsCOMT from metabolizing L-DOPA into 3-methoxy-4-hydroxy-L-phenylalaninein the periphery to avoid the undesirable effects of L-DOPA. The resultsshowed that entacapone increased efficacy of all L-Dopa and the pro-drugof L-Dopa treated groups (groups 1, 3, 4, 5, 6, 7, 9, 10), but did notchange the efficacy of vehicle group and pro-drug of aspirin treatedgroups (groups 2, 8 and 11).

When applying a combination of a plurality of drugs (e.g. one or moreHPPs and/or other drug(s)) to a subject, each drug could be appliedseparately, or one or more of the drugs could be applied atsubstantially the same time as separate drugs (e.g. spraying two or moredrugs at substantially the same time without mixing the drugs beforespraying), or one or more drugs could be mixed together before applyingto the subject, or any combination of the above application methods. Thedrugs could be applied in any order possible.

Example 38 The Efficacy of Drug A (Transdermally), Drug B(Transdermally), Entacapone (Orally) and Carbidopa Orally on Improvementof Motor Function Deficits and Reduction of Nigro-StriatalNeurodegeneration Induced by 6-OHDA in Parkinson's Disease (PD) Model atHDB

Test subjects (Sprague-Dawley rats) were prepared as described inExample 31, and grouped and tested following the protocol below:

-   1. Rats were divided into 11 groups (n=12), each group was applied    with the drug(s) at the doses specified in Table 9:

TABLE 9 Doses and Drug(s) Applied to the Test Animals Group GroupDrug(s) Parent Dosage Administra- No. Name Applied drug (mg/kg) tionMethod 1 Positive L-DOPA N/A 6 Oral Control Entacapone N/A 25 OralCarbidopa N/A 1.5 Oral 2 Negative 30% N/A 1,286 Transdermal Controlethanol(v/v) (μL/kg) (Vehicle) Entacapone N/A 25 Oral Carbidopa N/A 1.5Oral 3 Low dose Drug A L-DOPA 0.67 Transdermal Entacapone N/A 25 OralCarbidopa N/A 1.5 Oral 4 Moderate Drug A L-DOPA 2 Transdermal doseEntacapone N/A 25 Oral Carbidopa N/A 1.5 Oral 5 High dose Drug A L-DOPA6 Transdermal Entacapone N/A 25 Oral Carbidopa N/A 1.5 Oral 6 Drug AL-DOPA 0.67 Transdermal Drug B Aspirin 30 Transdermal Entacapone N/A 25Oral Carbidopa N/A 1.5 Oral 7 Drug A L-DOPA 2 Transdermal Drug B Aspirin30 Transdermal Entacapone N/A 25 Oral Carbidopa N/A 1.5 Oral 8 Drug BAspirin 30 Transdermal Entacapone N/A 25 Oral Carbidopa N/A 1.5 Oral 9Drug A L-DOPA 0.67 Transdermal Drug B Aspirin 90 Transdermal EntacaponeN/A 25 Oral Carbidopa N/A 1.5 Oral 10 Drug A L-DOPA 2 Transdermal Drug BAspirin 90 Transdermal Entacapone N/A 25 Oral Carbidopa N/A 1.5 Oral 11Drug B Aspirin 90 Transdermal Entacapone N/A 25 Oral Carbidopa N/A 1.5Oral

-   2. Dose Formulations-   (1) L-DOPA (3 mg/ml), entacapone (12.5 mg/ml), and carbidopa (1.5    mg/mL) in 0.5% CMC-Na was the positive control solution for group 1    (orally), the volume of administration was 2 mL/kg. Entacapone (12.5    mg/ml), and carbidopa (1.5 mg/mL) in 0.5% CMC-Na was the solution    for groups 2-11 (orally), the volume of administration was 2 mL/kg.    Vehicle solution (negative control solution for group 2) was 30%    ethanol (v/v), the volume of administration was 1,286 μL/kg. Other    test solutions (transdermally, groups 3-11) were freshly prepared    every day.-   (2) Preparation method of test solutions for groups 3-5: 50.33 mg of    Drug A was dissolved in 10 mL of 30% ethanol (v/v). This solution    was the stock solution (2).    -   a. The stock solution (2) was the test solution for Group 5        (high dose group) of Drug A. The volume of administration was        1286 μL/kg;    -   b. The test solution for group 4 (moderate dose group) of        Drug A. 3.00 mL of the stock solution (2) was diluted to final        volume 9.00 mL with 30% ethanol (v/v). This solution was the        test solution for Group 4. The volume of administration was 1286        μL/kg;    -   c. The test solution for group 3 (low dose group) of Drug A:        1.00 mL of the stock solution (2) was diluted to final volume        9.00 mL with 30% ethanol (v/v). This solution was the test        solution for Group 4. The volume of administration was 1286        μL/kg.-   (3) Preparation method of test solutions for group 6: 5.67 mg of    Drug A and 264 mg of Drug B were dissolved in 10 mL of 30% ethanol    (v/v). This solution was the test solution for groups 6;-   (4) Preparation method of test solutions for groups 7: 16.67 mg of    Drug A and 264 mg of Drug B were dissolved in 10 mL of 30% ethanol    (v/v). This solution was the test solution for groups 7;-   (5) Preparation method of test solutions for groups 8: 264 mg of    Drug B was dissolved in 10 mL of 30% ethanol (v/v). This solution    was the test solution for groups 8;-   (6) Preparation method of test solutions for group 9: 5.67 mg of    Drug A and 791 mg of Drug B were dissolved in 10 mL of 30% ethanol    (v/v). This solution was the test solution for groups 9;-   (7) Preparation method of test solutions for groups 10: 16.67 mg of    Drug A and 791 mg of Drug B were dissolved in 10 mL of 30% ethanol    (v/v). This solution was the test solution for groups 10;-   (8) Preparation method of test solutions for groups 11: 791 mg of    Drug B was dissolved in 10 mL of 30% ethanol (v/v). This solution    was the test solution for groups 11.-   3. On the day prior to the scheduled dosing, the hair (all rat    including the positive control group rats with L-DOPA due to the    double-blinded fashion) was removed from the up back (around the    neck and shoulder) of the animal using a small animal clipper. The    animals were re-clipped as necessary throughout the course of the    study to keep the epidermis exposed. On the day of dosing, an    aliquot (643 μL/kg) of the dosing formulation was administered to a    3 cm by 3 cm square section of the animal (around the neck and    shoulder) at around 9 am and repeat it at around 4 pm.-   4. At week 4 post lesion, the groups started to receive treatment.-   5. Rotarod test was performed 4 weeks after treatment.

FIG. 15 showed the staying time results in Totarod test described aboveafter 4 weeks treatment (n=12). FIG. 16 showed the falling speed resultsin Totarod test described above after 4 weeks treatment (n=12).

Applying a combination of carbidopa and entacapone worked better thanapplying either carbidopa or entacapone alone. The results showed thatthe combination of carbidopa and entacapone increased efficacy of allL-Dopa and the pro-drug of L-Dopa treated groups (groups 1, 3, 4, 5, 6,7, 9, 10), but did not change the efficacy of vehicle group and pro-drugof aspirin treated groups (groups 2, 8 and 11).

When applying a combination of a plurality of drugs (e.g. one or moreHPPs and/or other drug(s)) to a subject, each drug could be appliedseparately, or one or more of the drugs could be applied atsubstantially the same time as separate drugs (e.g. spraying two or moredrugs at substantially the same time without mixing the drugs beforespraying), or one or more drugs could be mixed together before applyingto the subject, or any combination of the above application methods. Thedrugs could be applied in any order possible.

Example 39 Treatment of Parkinson's Disease and Related Conditions

A solution of 15 mg of Drug A and 30 mg of Drug B in 0.5 ml of water isapplied (transdermally) to the skin on the neck, chest, back and anyother parts of a subject every 3-10 hours as need.

Example 40 Treatment of Parkinson's Disease and Related Conditions

A solution of 10 mg of Drug A and 20 mg of Drug B in 0.5 ml of water isapplied (transdermally) to the skin on the neck, chest, back and anyother parts of a subject every 3-10 hours as need.

Example 41 Treatment of Parkinson's Disease and Related Conditions

A solution of 20 mg of Drug A and 40 mg of Drug B in 0.5 ml of water isapplied (transdermally) to the skin on the neck, chest, back and anyother parts of a subject every 3-10 hours as need.

Example 42 Treatment of Parkinson's Disease and Related Conditions

A solution of 30 mg of Drug A and 50 mg of Drug B in 0.5 ml of water isapplied (transdermally) to the skin on the neck, chest, back and anyother parts of a subject every 3-10 hours as need.

Example 43 Treatment of Parkinson's Disease and Related Conditions

A solution of 15 mg of Drug A and 30 mg of Drug B in 0.5 ml of water isapplied (transdermally) to the skin on the neck, chest, back and/or anyother parts of a subject every 3-10 hours as need. After or beforeapplying the above solution, a tablet containing 10 mg of carbidopaand/or inactive ingredients, such as, cellulose, mannitol,croscarmellose sodium, vegetable oil, hydroxypropyl methylcellulose, istaken orally to increase the efficacy of the transdermally treatment.

Example 44 Treatment of Parkinson's Disease and Related Conditions

A solution of 20 mg of Drug A and 40 mg of Drug B in 0.5 ml of water isapplied (transdermally) to the skin on the neck, chest, back and/or anyother parts of a subject every 3-10 hours as need. After or beforeapplying the above solution, a tablet containing 15 mg of carbidopaand/or inactive ingredients, such as, cellulose, mannitol,croscarmellose sodium, vegetable oil, hydroxypropyl methylcellulose, istaken orally to increase the efficacy of the transdermally treatment.

Example 45 Treatment of Parkinson's Disease and Related Conditions

A solution of 15 mg of Drug A and 30 mg of Drug B in 0.5 ml of water isapplied (transdermally) to the skin on the neck, chest, back and/or anyother parts of a subject every 3-10 hours. After or before applying theabove solution, a tablet containing 15 mg of carbidopa, 70 mg ofentacapone and/or inactive ingredients, such as, cellulose, mannitol,croscarmellose sodium, vegetable oil, hydroxypropyl methylcellulose, istaken orally to increase the efficacy of the transdermally treatment.

Example 46 Treatment of Parkinson's Disease and Related Conditions

A solution of 20 mg of Drug A and 40 mg of Drug B in 0.5 ml of water isapplied (transdermally) to the skin on the neck, chest, back and/or anyother parts of a subject every 3-10 hours. After or before applying theabove solution, a tablet containing 15 mg of carbidopa, 100 mg ofentacapone and/or inactive ingredients, such as, cellulose, mannitol,croscarmellose sodium, vegetable oil, hydroxypropyl methylcellulose, istaken orally to increase the efficacy of the transdermally treatment.

Example 47 Treatment of Parkinson's Disease and Related Conditions

A solution of 10 mg of Drug C and 15 mg of Drug D in 0.5 ml of water isapplied (transdermally) to the skin on the neck, chest, back and/or anyother parts of a subject every 3-10 hours.

Example 49 Treatment of Parkinson's Disease and Related Conditions

A solution of 30 mg of Drug C and 30 mg of Drug D in 0.5 ml of water isapplied (transdermally) to the skin on the neck, chest, back and/or anyother parts of a subject every 3-10 hours.

Example 50 Treatment of Parkinson's Disease and Related Conditions

A solution of 20 mg of Drug C and 15 mg of Drug D in 0.5 ml of water isapplied (transdermally) to the skin on the neck, chest, back and/or anyother parts of a subject every 3-10 hours. After or before applying theabove solution, a tablet containing 15 mg of carbidopa and/or inactiveingredients, such as, cellulose, mannitol, croscarmellose sodium,vegetable oil, hydroxypropyl methylcellulose, is taken orally toincrease the efficacy of the transdermally treatment.

Example 51 Treatment of Parkinson's Disease and Related Conditions

A solution of 10 mg of Drug C and 10 mg of Drug D in 0.5 ml of water isapplied (transdermally) to the skin on the neck, chest, back and/or anyother parts of a subject every 3-10 hours. After or before applying theabove solution, a tablet containing 15 mg of carbidopa, 100 mg ofentacapone and/or inactive ingredients, such as, cellulose, mannitol,croscarmellose sodium, vegetable oil, hydroxypropyl methylcellulose, istaken orally to increase the efficacy of the transdermally treatment.

Example 52 Treatment of Parkinson's Disease and Related Conditions

A solution of 30 mg of Drug C and 20 mg of Drug D in 0.5 ml of water isapplied (transdermally) to the skin on the neck, chest, back and/or anyother parts of a subject every 3-10 hours. After or before applying theabove solution, a tablet containing 15 mg of carbidopa, 70 mg ofentacapone and/or inactive ingredients, such as, cellulose, mannitol,croscarmellose sodium, vegetable oil, hydroxypropyl methylcellulose, istaken orally to increase the efficacy of the transdermally treatment.

Example 53 Treatment of Parkinson's Disease and Related Conditions

A solution of 30 mg of Drug C and 25 mg of Drug D in 0.5 ml of water isapplied (transdermally) to the skin on the neck, chest, back and/or anyother parts of a subject every 3-10 hours. After or before applying theabove solution, a tablet containing 10 mg of carbidopa, 50 mg ofentacapone and/or inactive ingredients, such as, cellulose, mannitol,croscarmellose sodium, vegetable oil, hydroxypropyl methylcellulose, istaken orally to increase the efficacy of the transdermally treatment.

Example 54 Treatment of Parkinson's Disease and Related Conditions

A solution of 10 mg of Drug E and 10 mg of Drug F in 0.5 ml of water isapplied (transdermally) to the skin on the neck, chest, back and/or anyother parts of a subject every 3-10 hours.

Example 55 Treatment of Parkinson's Disease and Related Conditions

A solution of 20 mg of Drug E and 20 mg of Drug F in 0.5 ml of water isapplied (transdermally) to the skin on the neck, chest, back and/or anyother parts of a subject every 3-10 hours.

Example 56 Treatment of Parkinson's Disease and Related Conditions

A solution of 5 mg of Drug E and 10 mg of Drug F in 0.5 ml of water isapplied (transdermally) to the skin on the neck, chest, back and/or anyother parts of a subject every 3-10 hours.

Example 57 Treatment of Parkinson's Disease and Related Conditions

A solution of 10 mg of Drug G and 10 mg of Drug H in 0.5 ml of water isapplied (transdermally) to the skin on the neck, chest, back and/or anyother parts of a subject every 3-10 hours.

Example 58 Treatment of Parkinson's Disease and Related Conditions

A solution of 15 mg of Drug G and 15 mg of Drug H in 0.5 ml of water isapplied (transdermally) to the skin on the neck, chest, back and/or anyother parts of a subject every 3-10 hours.

Example 59 Treatment of Parkinson's Disease and Related Conditions

A solution of 20 mg of Drug G and 15 mg of Drug H in 0.5 ml of water isapplied (transdermally) to the skin on the neck, chest, back and/or anyother parts of a subject every 3-10 hours.

Example 60 Treatment of Parkinson's Disease and Related Conditions

A solution of 20 mg of Drug G and 15 mg of Drug H in 0.5 ml of water isapplied (transdermally) to the skin on the neck, chest, back and/or anyother parts of a subject every 3-10 hours. After or before applying theabove solution, a tablet containing 10 mg of carbidopa, 70 mg ofentacapone and/or inactive ingredients, such as, cellulose, mannitol,croscarmellose sodium, vegetable oil, hydroxypropyl methylcellulose, istaken orally to increase the efficacy of the transdermally treatment.

Example 61 Treatment of Parkinson's Disease and Related Conditions

A solution of 10 mg of Drug G and 10 mg of Drug H in 0.5 ml of water isapplied (transdermally) to the skin on the neck, chest, back and/or anyother parts of a subject every 3-10 hours. After or before applying theabove solution, a tablet containing 10 mg of carbidopa, 50 mg ofentacapone and/or inactive ingredients, such as, cellulose, mannitol,croscarmellose sodium, vegetable oil, hydroxypropyl methylcellulose, istaken orally to increase the efficacy of the transdermally treatment.

Example 62 Treatment of Parkinson's Disease and Related Conditions

A solution of 20 mg of Drug G and 15 mg of Drug H in 0.5 ml of water isapplied (transdermally) to the skin on the neck, chest, back and/or anyother parts of a subject every 3-10 hours. After or before applying theabove solution, a tablet containing 15 mg of carbidopa, 100 mg ofentacapone and/or inactive ingredients, such as, cellulose, mannitol,croscarmellose sodium, vegetable oil, hydroxypropyl methylcellulose, istaken orally to increase the efficacy of the transdermally treatment.

What is claimed is:
 1. A method for treating Parkinson disease in asubject comprising administering transdermally a pharmaceuticalcomposition comprising one or more high penetration pro-drugs selectedfrom Structure Pro-L-Dopa-1, Structure Pro-L-Dopa-2, StructurePro-L-Dopa-3, Structure Pro-L-Dopa-4, Structure Pro-L-Dopa-5, StructurePro-dopamine-1, Structure Pro-dopamine-2, Structure Pro-dopamine-3,Structure Pro-dopamine-4, and Structure Pro-dopamine-5:

stereoisomers, and pharmaceutically acceptable salts thereof, wherein Wis selected from H, substituted and unsubstituted alkyl, substituted andunsubstituted cycloalkyl, substituted and unsubstitutedheterocycloalkyl, substituted and unsubstituted alkyloxy, substitutedand unsubstituted alkenyl, substituted and unsubstituted alkynyl,substituted and unsubstituted aryl, substituted and unsubstitutedheteroaryl, the protonatable amine group, pharmaceutically acceptablesubstituted and unsubstituted amine groups, Structure W-1, StructureW-2, Structure W-3, Structure W-4, Structure W-5, Structure W-6,Structure W-7, Structure W-8, Structure W-9, Structure W-10, StructureW-11, and Structure W-12:

HA is selected from nothing and a pharmaceutically acceptable acid; X isselected from O, S, and NR₃; X₂ is selected from nothing, O, S, NR₃,CHR₃—O, CHR₃—S, CHR₃—O, O—CHR₃—O, O—CHR₃—S, S—CHR₃—O, and S—CHR₃—S; X₃is selected from nothing, C═O, C═S, C(═O)—O, O, S, NR₃, C(═O)—O—CHR₃—O,C(═O)—O—CHR₃—S, C(═O)—S—CHR₃—O, and C(═O)—S—CHR₃—S; Y₁ is selected fromR₃C(═O), R₃O—C(═O), and R₃S—C(═O); Y₂ is selected from R₃C(═O),R₃O—C(═O), and R₃S—C(═O); Y₃ is selected from R₃, OR₃, SR₃, N R₃R₄,O—CHR₃—OR₄, O—CHR₃—SR₄, S—CHR₃—OR₄; n and m are independently selectedfrom 0, 1, 2, 3, 4, 5, 6, 7, and 8; Rc is selected from nothing,CH₂C(═O)OR₆, substituted and unsubstituted alkyl, substituted andunsubstituted cycloalkyl, substituted and unsubstitutedheterocycloalkyl, substituted and unsubstituted alkoxyl, substituted andunsubstituted perfluoroalkyl, substituted and unsubstituted alkylhalide, substituted and unsubstituted alkenyl, substituted andunsubstituted alkynyl, substituted and unsubstituted aryl, andsubstituted and unsubstituted heteroaryl, wherein any CH₂ in R may befurther replaced with O, S, P, NR₆, or any other pharmaceuticallyacceptable groups; R is selected from nothing, CH₂C(═O)OR₆, substitutedand unsubstituted alkyl, substituted and unsubstituted cycloalkyl,substituted and unsubstituted heterocycloalkyl, substituted andunsubstituted alkoxyl, substituted and unsubstituted perfluoroalkyl,substituted and unsubstituted alkyl halide, substituted andunsubstituted alkenyl, substituted and unsubstituted alkynyl,substituted and unsubstituted aryl, and substituted and unsubstitutedheteroaryl, wherein any CH₂ in R may be further replaced with O, S, P,NR₆, or any other pharmaceutically acceptable groups; R₃ is selectedfrom nothing, CH₂C(═O)OR₆, substituted and unsubstituted alkyl,substituted and unsubstituted cycloalkyl, substituted and unsubstitutedheterocycloalkyl, substituted and unsubstituted alkoxyl, substituted andunsubstituted perfluoroalkyl, substituted and unsubstituted alkylhalide, substituted and unsubstituted alkenyl, substituted andunsubstituted alkynyl, substituted and unsubstituted aryl, andsubstituted and unsubstituted heteroaryl, wherein any CH₂ in R may befurther replaced with O, S, P, NR₆, or any other pharmaceuticallyacceptable groups; each R₆ is independently selected from H, F, Cl, Br,I, Na⁺, K⁺, C(═O)R₅, 2-oxo-1-imidazolidinyl, phenyl, 5-indanyl,2,3-dihydro-1H-inden-5-yl, 4-hydroxy-1,5-naphthyridin-3-yl, substitutedand unsubstituted alkyl, substituted and unsubstituted cycloalkyl,substituted and unsubstituted heterocycloalkyl, substituted andunsubstituted alkenyl, substituted and unsubstituted alkynyl,substituted and unsubstituted alkyloxyl, substituted and unsubstitutedcycloalkyloxyl, substituted and unsubstituted aryl, and substituted andunsubstituted heteroaryl; each R₅ is independently selected from H,C(═O)NH₂, CH₂CH₂N(CH₃)₂, CH₂CH₂N(CH₂CH₃)₂, Cl, F, Br, I, substituted andunsubstituted alkyl, substituted and unsubstituted cycloalkyl,substituted and unsubstituted heterocycloalkyl, substituted andunsubstituted alkyloxyl, substituted and unsubstituted cycloalkyloxyl,substituted and unsubstituted aryl, substituted and unsubstitutedheteroaryl, substituted and unsubstituted alkylcarbonyl, and substitutedand unsubstituted alkylamino; R₁ and R₂ are independently selected fromH, substituted and unsubstituted alkyl, substituted and unsubstitutedcycloalkyl, substituted and unsubstituted heterocycloalkyl, substitutedand unsubstituted alkyloxyl, substituted and unsubstituted alkenyl,substituted and unsubstituted alkynyl, substituted and unsubstitutedaryl and substituted and unsubstituted heteroaryl residues; R₁₁-R₁₅ areindependently selected from nothing, H, substituted and unsubstitutedalkyl, substituted and unsubstituted cycloalkyl, substituted andunsubstituted heterocycloalkyl, substituted and unsubstituted alkoxyl,substituted and unsubstituted perfluoroalkyl, substituted andunsubstituted alkyl halide, substituted and unsubstituted alkenyl,substituted and unsubstituted alkynyl, substituted and unsubstitutedaryl, and substituted and unsubstituted heteroaryl; and an agentselected from a high penetration prodrug of aspirin, a high penetrationprodrug of another nonsteroidal anti-inflammatory drug (NSAID), and anycombination thereof; and administering orally an aromatic-L-amino-aciddecarboxylase inhibitor selected from carbidopa, benserazide,difluromethyldopa, α-methyldopa, and a pharmaceutical compositionthereof; and/or administering a catechol-O-methyl transferase inhibitorselected from entacapone, tolcapone; and a pharmaceutical compositionthereof.
 2. A method for treating Parkinson disease in a subjectcomprising administering transdermally a pharmaceutical compositioncomprising 4-(2-(2-(methylamino)acetamido)ethyl)-1,2,-phenylenedibenzoate hydrochloride,4-(2-(2-(methylamino)acetamido)ethyl)-1,2,-phenylene dibenzoate aceticacid,4-(6-methyl-4,8-dioxo-5,7-dioxa-2,9-diazadecan-11-yl)-1,2,-phenylenedibenzoate hydrobromide,4-(2-(2-amino-3-phenylpropanamido)ethyl)-1,2,-phenylene dibenzoatehydrochloride,4-(2-(((1-((pyrrolidine-2-carbonyl)oxy)ethoxy)amino)ethyl)-1,2,-phenylenedibenzoate hydrochloride,4-(2-piperidine-4-carboxamido)ethyl)-1,2,-phenylenebis(2-ethylbutanoate) hydrochloride,4-(2-((((-octahydro-1H-quinolizin-3-yl)oxy)carbonyl)amino)ethyl)-1,2,-phenylenebis(2-ethylbutanoate) acetate,1-(((2-(4-amino-2,5-dioxo-2,3,4,5-tetrahydrobenzo[b][1,4]dioxocin-8-yl)ethyl)carbamoyl)oxy)ethylisobutyrate hydrofluoride,1-(((2-(3-amino-2,5-dioxo-2,3,4,5-tetrahydrobenzo[b][1,4]dioxocin-8-yl)ethyl)carbamoyl)oxy)ethylisobutyrate hydrofluoride,(-(2-((ethoxycarbonyl)amino)ethyl)-2-(2-methylamino)acetoxy)phenylbenzoate hydrochloride,5-(2-((ethoxycarbonyl)amino)ethyl)-2-(2-methylamino)acetoxy)phenylbenzoate hydrochloride, 4-(2-aminoethyl)-1,2-phenylene dibenzoatehydrochloride, (S)-4-(2-amino-3-isopropoxy-3-oxopropyl)-1,2-phenylenedibenzoate hydrochloride,(S)-4-(2-amino-3-(heptan-4-yloxy)-3-oxopropyl)-1,2-phenylene dibenzoatehydrochloride, (S)-4-(2-amino-3-isopropoxy-3-oxopropyl)-1,2-phenylenedipentanoate hydrochloride,(S)-4-(2-amino-3-ethoxy-3-oxopropyl)-1,2-phenylene diacetatehydrochloride,(S)-4-(2-amino-3-oxo-3-(pentan-3-yloxy)propyl)-1,2-phenylenebis(2-methylpropanoate) hydrobromide,(S)-4-(2-aminoacetamido)-3-isopropoxy-3-oxopropyl)-1,2-phenylenedibenzoate hydrochloride,4-((2S)-3-oxo-3-(pentan-3-yloxy)-2-(pyrrolidine-2-carboxamido)propyl)-1,2-phenylenedibenzoate hydrofluoride,(S)-4-(3-isopropoxy-3-oxo-2-(piperidine-4-carboxamido)propyl)-1,2-phenylenedibenzoate hydrochloride,4-((2S)-3-isopropoxy-3-2-(octahydro-1H-quinolizine-2-carboxamido)-3-oxopropyl)-1,2-phenylenebis(2-methylpropanoate) hydrochloride, (2S)-isopropyl3-(3-amino-2,5-dioxo-2,3,4,5-tetrahydrobenzo[b][1,4]dioxocin-8-yl)-2-(((1-(isobutyryloxy)ethoxy)carbonyl)amino)propanoatehydrobromide, (2S)-isopropyl3-(4-amino-2,5-dioxo-2,3,4,5-tetrahydrobenzo[b][1,4]dioxocin-8-yl)-2-(((1-(isobutyryloxy)ethoxy)carbonyl)amino)propanoatehydrobromide,5-((2S)-2-(((1-(isobutyryloxy)ethoxy)carbonyl)amino-3-isopropoxy-3-oxopropyl)-2-(2-(methylamino)acetoxy)phenylbenzoate hydrochloride,4-((2S)-2-(((1-(isobutyryloxy)ethoxy)carbonyl)amino-3-isopropoxy-3-oxopropyl)-2-(2-(methylamino)acetoxy)phenylbenzoate hydrochloride,4-((2S)-3-isopropoxy-2-((((octahydroindolizin-1-yl)oxy)carbonyl)amino)-3-oxopropyl)-1,2-phenylenedibenzoate acetate, 2-(diethylamino)ethyl2-[(2,6-dichloro-3-methylphenyl)amino]benzoate.acetate,(Z)-2-(diethylaminoethyl)ethyl 2-(5-fluoro-2-methyl-1-(4-methylsulfinyl)benzylidene)-1H-inden-1-yl)acetate.AcOH, 2-(dimethylamino)ethyl2-(3-phenoxyphenyl) propionate.hydrochloride, S-(2-(dimethylamino)ethyl2-(3-phenoxyphenyl) propanethioate hydrochloride, 2-(dipropylamino)ethyl4-acetoxy-2′,4′-difluoro-[1,1′-biphenyl]-3-carboxylate hydrochloride,2-(diethylamino)ethyl 2-(4-isobutylphenyl)propionate hydrochloride,and/or 2-(diethylamino)ethyl 2-acetoxybenzoate hydrochloride, and/or apharmaceutically acceptable salt thereof; and administering orally anaromatic-L-amino-acid decarboxylase inhibitor selected from carbidopa,benserazide, difluromethyldopa, α-methyldopa, and a pharmaceuticalcomposition thereof; and/or administering a catechol-O-methyltransferase inhibitor selected from entacapone, tolcapone, and apharmaceutical composition thereof.
 3. The method of claim 2 comprisingadministering transdermally a pharmaceutical composition comprising(S)-4-(2-amino-3-isopropoxy-3-oxopropyl)-1,2-phenylene dibenzoate and2-(diethylamino)ethyl 2-acetoxybenzoate, and/or pharmaceuticallyacceptable salt thereof; and administering orally anaromatic-L-amino-acid decarboxylase inhibitor selected from carbidopa,benserazide, difluromethyldopa, α-methyldopa, and a pharmaceuticalcomposition thereof; and/or administering a catechol-O-methyltransferase inhibitor selected from entacapone, tolcapone; and apharmaceutical composition thereof.
 4. The method of claim 2 comprisingadministering transdermally a pharmaceutical composition comprising(S)-4-(2-amino-3-isopropoxy-3-oxopropyl)-1,2-phenylene dibenzoate,2-(diethylamino)ethyl 2-acetoxybenzoate, carbidopa and/or entacapone,and/or pharmaceutically acceptable salt thereof; and administeringorally an aromatic-L-amino-acid decarboxylase inhibitor selected fromcarbidopa, benserazide, difluromethyldopa, α-methyldopa, and apharmaceutical composition thereof; and/or administering acatechol-O-methyl transferase inhibitor selected from entacapone,tolcapone; and a pharmaceutical composition thereof.
 5. The method ofclaim 2 comprising administering transdermally a pharmaceuticalcomposition comprising(S)-4-(2-amino-3-oxo-3-(pentan-3-yloxy)propyl)-1,2-phenylenebis(2-methylpropanoate) and 2-(diethylamino)ethyl2-(4-isobutylphenyl)propionate, and/or pharmaceutically acceptable saltthereof; and administering orally an aromatic-L-amino-acid decarboxylaseinhibitor selected from carbidopa, benserazide, difluromethyldopa,α-methyldopa, and a pharmaceutical composition thereof; and/oradministering a catechol-O-methyl transferase inhibitor selected fromentacapone, tolcapone; and a pharmaceutical composition thereof.
 6. Themethod of claim 2 comprising administering transdermally apharmaceutical composition comprising4-(2-(((1-((pyrrolidine-2-carbonyl)oxy)ethoxy)amino)ethyl)-1,2,-phenylenedibenzoate and 4-(dimethylamino)butyl 2-(3-phenoxyphenyl) propionate,and/or pharmaceutically acceptable salt thereof; and administeringorally an aromatic-L-amino-acid decarboxylase inhibitor selected fromcarbidopa, benserazide, difluromethyldopa, α-methyldopa, and apharmaceutical composition thereof; and/or administering acatechol-O-methyl transferase inhibitor selected from entacapone,tolcapone; and a pharmaceutical composition thereof.
 7. The method ofclaim 2 comprising administering transdermally a pharmaceuticalcomposition comprising(S)-4-(2-amino-3-oxo-3-(pentan-3-yloxy)propyl)-1,2-phenylene,bis(2-methylpropanoate), 2-(diethylamino)ethyl2-(4-isobutylphenyl)propionate, carbidopa and/or entacapone, and/orpharmaceutically acceptable salt thereof; and administering orally anaromatic-L-amino-acid decarboxylase inhibitor selected from carbidopa,benserazide, difluromethyldopa, α-methyldopa, and a pharmaceuticalcomposition thereof; and/or administering a catechol-O-methyltransferase inhibitor selected from entacapone, tolcapone; and apharmaceutical composition thereof.
 8. The method of claim 2 comprisingadministering transdermally a pharmaceutical composition comprising(S)-4-(2-amino-3-(heptan-4-yloxy)-3-oxopropyl)-1,2-phenylene dibenzoate,and 2-(dipropylamino)ethyl4-acetoxy-2′,4′-difluoro-[1,1′-biphenyl]-3-carboxylate, and/orpharmaceutically acceptable salt thereof; and administering orally anaromatic-L-amino-acid decarboxylase inhibitor selected from carbidopa,benserazide, difluromethyldopa, α-methyldopa, and a pharmaceuticalcomposition thereof; and/or administering a catechol-O-methyltransferase inhibitor selected from entacapone, tolcapone; and apharmaceutical composition thereof.
 9. The method of claim 2 comprisingadministering transdermally a pharmaceutical composition comprising(S)-4-(2-amino-3-(heptan-4-yloxy)-3-oxopropyl)-1,2-phenylene dibenzoate,and 2-(dipropylamino)ethyl4-acetoxy-2′,4′-difluoro-[1,1′-biphenyl]-3-carboxylate, carbidopa and/orentacapone, and/or pharmaceutically acceptable salt thereof; andadministering orally an aromatic-L-amino-acid decarboxylase inhibitorselected from carbidopa, benserazide, difluromethyldopa, α-methyldopa,and a pharmaceutical composition thereof; and/or administering acatechol-O-methyl transferase inhibitor selected from entacapone,tolcapone; and a pharmaceutical composition thereof.